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Abstract Ethnopharmacological relevance Respiratory diseases and asthma, in particular, are nowadays a global health problem. In Rwanda, some traditional healers claim to treat asthma with plant-based recipes, though there is no scientific proof so far. Aim of the study Our study aimed at evaluating the toxicity and the anti-inflammatory effect of plant recipes used in Rwanda against asthma in order to select potential candidates for further characterization of the active compounds. Materials and methods Water (aqueous) and methanol-dichloromethane (organic) extracts from selected folkloric recipes were submitted for toxicity test on THP-1 derived macrophages using CellTiter-Glo Luminescent Cell Viability Assay. The evaluation of the anti-inflammatory effect of the plant extracts was carried out using the Caspase-Glo 1 Inflammasome assay on THP-1 -derived macrophages. Results Most of both organic and aqueous extract showed more than 95% of cell viability up to 200 µg/ml, except for R03Cn organic extract that inhibited 25% of the cell viability. Plant extracts inhibited caspase-1 activation in THP-1 derived macrophages in a dose-dependent manner. Four extracts (R03Cn and R07Kn aqueous extracts, R10MK and R19Sz organic extracts) strongly downregulated the activation of caspase-1 (more than 70% at 50 µg/ml). In general, organic extracts exhibited better caspase-1 inhibitory effects than their aqueous counterparts. Conclusions The inhibition of inflammasome/caspase-1 is one of key mechanisms of action in asthma. Some traditional recipes are active on this mechanism and are thus strong candidates for the treatment of asthma and other inflammasome-mediated diseases. Further investigations are needed to characterize active molecules. Graphical abstract fx1 [ABSTRACT FROM AUTHOR]

Abstract Ethnopharmacological relevance Acanthus mollis is a plant native to the Mediterranean region, traditionally used as diuretic, anti-inflammatory and soothing of the mucous membranes of the digestive and urinary tract and externally as healing of wounds and burns, also demonstrating analgesic and anti-inflammatory activities. However, studies focused on its phytochemical composition as well as scientific proof of Acanthus mollis efficacy are scarce. Aim of the study The proposed work aims to perform a phytochemical characterization and evaluation of the therapeutic potential of Acanthus mollis , based on biological properties that support its traditional uses. Material and methods In this study, an 96% ethanol extract from Acanthus mollis leaves was obtained and its phytochemical composition evaluated using High Performance Liquid Chromatography with Photodiode Array Detector coupled to Electrospray Ionization Mass Spectrometry (HPLC-PDA-ESI/MSn). The chemical structure of the compound isolated was elucidated using 1H and 13C Nuclear Magnetic Resonance (NMR), 1H-correlation spectroscopy (1H–COSY), heteronuclear single quantum correlation (HSQC) and heteronuclear multiple-bond correlation (HMBC). The quantification of the constituents was performed using two external standards (2,4-dihydroxy-1,4-benzoxazin-3-one and verbascoside). The antioxidant activity was determined by the 2,2-diphenyl-1-pycrylhydrazyl (DPPH) assay. Anti-inflammatory activity was determined measuring the inhibition of nitric oxide production by RAW 264.7 macrophages stimulated with the TLR4 agonist lipopolysaccharide (LPS) and through lipoxygenase (LOX) inhibition assay. The cytotoxicity was screened on two lines (RAW 264.7 and HaCaT) using the resazurin assay. Results Compounds such as verbascoside and its derivatives, as well as benzoxazinoids were found as the main constituents. A percentage of 5.58% was verified for the 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) derivatives. DIBOA was the main compound of the extract. Significant concentrations were also found for phenylpropanoids, which constitute about 4.39% of the total compounds identified. This extract showed antioxidant capacity against DPPH (IC 50 = 40.00 ± 1.59 μg/mL) and superoxide anion (IC 50 = 29.42 ± 1.99 μg/mL). It also evidenced anti-inflammatory potential in RAW 264.7 macrophages, presenting capacity for nitric oxide reduction (IC 50 = 28.01 μg/mL). Moreover, in vitro studies have shown that this extract was able to inhibit the lipoxygenase, with an IC 50 of 104.39 ± 4.95 µg/mL. Importantly, all effective concentrations were devoid of cytotoxicity in keratinocytes, thus highlighting the safety of the extract for the treatment of skin inflammatory related diseases. Concerning macrophages it was also possible to disclose concentrations showing anti-inflammatory activity and without cytotoxicity (up to 30 µg/mL). The benzoxazinoid DIBOA demonstrated a considerable anti-inflammatory activity suggesting its important contribution to this activity. Conclusions These results corroborate the anti-inflammatory properties traditionally attributed to this plant. Among the compounds identified in this study, benzoxazinoids exhibited a significant anti-inflammatory activity that was never previously described. Ethanol seems to be a good option for the extraction of these bioactive compounds, since relevant antioxidant/anti-radical and anti-inflammatory activities were found for this extract. Graphical abstract fx1 [ABSTRACT FROM AUTHOR]

Abstract Ethnopharmacological relevance Tuberculosis (TB) is one of the highly infectious disease caused by the bacterium Mycobacterium tuberculosis (Mtb). Several medicinal plants are traditionally used by the tribal healers of some tribal pockets of Madhya Pradesh (M.P.), India in the treatment of various ailments including TB related symptoms. The information of traditional knowledge of plants is empirical lacking systematic scientific investigation. Aim of the study To provide a scientific rationale for the traditional uses of some medicinal plants in treating prolonged cough, chest complaints and TB by ethnic groups in four districts of the state of M.P., India. The anti-mycobacterial activity of these plants was also investigated against multidrug-resistant (MDR) strains of Mtb. The active plant extracts were also assessed for general cytotoxicity by human monocytic leukemia cell line, THP-1 macrophages. Materials and methods Semi structured interviews and guided field-walk methods were used to gather information on medicinal plants used by the tribal traditional healers. The study was carried out in the tribal pockets of Anuppur, Mandla, Umariya and Dindori districts of M.P., India. A total of 35 plant species distributed in 22 families were selected based on folklore uses suggested by the tribal traditional healer. The ethanol extracts of plants were evaluated for anti-mycobacterial activity against Mtb H37Rv and six MDR clinical isolates of Mtb. The anti-mycobacterial activity of plants was determined in terms of minimum inhibitory concentration (MIC) using standard resazurin microtitre plate assay (REMA). The plant extracts found to be active against Mtb were further evaluated for general cytotoxicity against human THP-1 macrophages using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. The selectivity index (SI) of active plant extracts were also calculated on the basis of MIC and cytotoxicity. Results Out of the 35 plants, 11 plant species showed anti-mycobacterial activity with MIC ranging from 500 to 31.25 µg/mL against Mtb H37Rv. The plant extracts also exhibited anti-mycobacterial activity against six multi-drug resistant (MDR) clinical strains of Mtb isolated from sputum samples of pulmonary TB patients. In vitro cytotoxicity of active plant extracts was also assessed in human THP-1 macrophages. The IC 50 (50% inhibitory concentration) values of most of the plant extracts on THP-1 was found to be higher than MIC values against Mtb , indicating that the THP-1 cells are not adversely affected at concentrations that are effective against Mtb. Significant anti-mycobacterial activity was observed for eleven plants viz., Alstonia scholaris (L.) R. Br. , Glycyrrhiza glabra L. , Holorrhena antidysentrica (Roth) Wall. exA.DC. , Mallotus philippensis (Lam.) Müll. Arg. , Eulophia nuda Lindl. , Cocculus hirsutus (L.) Diels , Pueraria tuberosa (Willd.) DC. , Cyperus rotundus L. , Curcuma caesia Roxb. , Sphaeranthus indicus L. and Plumbago zeylanica L. which lends support to their traditional uses. Conclusion The present investigation supports the potential role of plants used by tribal healers as our results have shown that these plants exhibit anti-mycobacterial activity in the acceptable range against Mtb. Our study clearly lends support to the traditional uses of some plants in TB related symptoms as we have found them to exhibit significant in vitro anti-TB activity. To the best of our knowledge, anti-mycobacterial activity in plants against Mtb is being reported for the first time from tribal areas of M.P., India by the current study. Graphical abstract fx1 [ABSTRACT FROM AUTHOR]

Abstract Ethnopharmacological relevance Capsicum annum L. (CA) is used extensively as a spice and is a rich source of antioxidant vitamins. It has long been used in Indian, Native American, and Chinese traditional medicine as a carminative and an appetizer that normalizes liver function. However, its hepato-protective activity has so far not been studied. Aim of the study The present study was undertaken to evaluate the efficacy of aqueous extract of CA at two different doses (125 mg/kg body weight and 250 mg/kg body weight), against ethanol induced oxidative stress and apoptosis in liver tissue. Materials and methods Adult male Wistar rats, weighing 150–200 g, were randomly grouped (n = 6) and treated with ethanol (2 g/kg bw, i.p.), CA 125 (125 mg/kg bw, i.p.), CA 250 (250 mg/kg bw, i.p.), ethanol with CA (similar doses), and control (0.5 ml normal saline, i.p.) for 30 days. Lipid peroxidation (LPO) and reduced glutathione content (GSH) in tissue homogenate, along with catalase (CAT), superoxide dismutase (Cu-Zn-SOD & Mn-SOD), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-s-transferase (GST) and glucose-6-phosphate dehydrogenase (G-6-P-D) activity were evaluated. Serum levels of alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphate (ALP), triglyceride (TG), total cholesterol (CHLS), high density lipoprotein (HDL), low density lipoprotein (LDL) very low density lipoprotein (VLDL), tumour necrotic factor alpha (TNF-α) and interleukin 6 (IL-6) were also measured using ELISA kits. Histopathological evaluation of the hepatic tissue was performed by hematoxylin and eosin (H&E) and periodic acid-schiff (PAS) staining. TUNEL assay was performed for apoptosis detection. Results Ethanol significantly (p < 0.001) increased ALT, AST, ALP, TNF-α, IL-6, LPO, Cu-Zn-SOD, GST, GPx, TG, CHLS, LDL, VLDL levels, along with significant (p < 0.001) decrease in HDL, Mn-SOD, CAT, GSH, GR and G6PD activity. Co-administration of CA along with ethanol alleviated changes in the above parameters (p < 0.001) in a dose-dependent manner and also reduced the number of apoptotic death cells. Histo-pathological and histo-chemical studies of liver sections also ascertained the outcomes of this study. Conclusion Thus, it can be concluded that the aqueous extract of green CA can exert a protective effect against ethanol induced hepato-toxicity. The possible mechanism may be by acting as an antioxidant; preventing ethanol induced apoptosis and reducing pro-inflammatory cytokine levels. Graphical abstract fx1 Highlights • Alcohol causes hepatic damage by the way of generating oxidative stress. • Capsicum annum (CA) is a commonly used spice worldwide. • Extract of green CA was studied as a hepatoprotective agent against ethanol. • Aqueous extract of CA can ameliorate the hepato-cellular damage caused by ethanol. • CA prevents fibrosis, DNA-fragmentation and cell apoptosis. [ABSTRACT FROM AUTHOR]

A new bakuchiol compound Δ 11 -12-hydroxy-12-dimethyl bakuchiol ( 1 ), a new flavanone compound 2( S )-6-methoxy-7- hydroxymethylene-4′-hydroxyl-flavanone ( 8 ), and two new isoflavanone compounds 4′,7-dihydroxy-3′-(6”β-hydroxy-3″,7″-dimethyl-,2″,7″-dibutenyl)-geranylisoflavone ( 9 ) and 4′,7-dihydroxy-3′-(7″-hydroxy-7″-methyl-2″,5″-dibutenyl)-geranylisoflavone ( 10 ) together with eight known compounds ( 2–7 , 11 , 12 ) were isolated from the P. corylifolia . Their structures were elucidated on the basis of spectroscopic and physico-chemical analyses. All the isolates were evaluated for in vitro inhibitory activity against DGAT1/2. Among them, compounds 3 , 9 and 10 were found to exhibit selective inhibitory activity on DGAT1 with IC 50 values ranging from 93.7 ± 1.3 to 96.2 ± 1.1 μM. Compound 1 showed inhibition activity on DGAT1 with IC 50 values 73.4 ± 1.3 μM and inhibition of DGAT2 with IC 50 value 121.1 ± 1.3 μM. [ABSTRACT FROM AUTHOR]

Four new cinnamoyl-phloroglucinols ( 1 – 4 ) were isolated from the leaves of Xanthostemon chrysanthus . Compounds 1 and 2 represent the first example of natural phloroglucinols with an oxazole unit. Their structures were elucidated on the basis of NMR spectroscopic data and single crystal X-ray diffraction. Compound 3 showed moderate cytotoxic activity against MDA-MB-231 and SGC-7901 cells with IC 50 values of 25.26 ± 0.35 μM and 31.2 ± 0.94 μM, respectively. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Ginger ( Zingiber officinale Roscoe) is a well known anti-inflammatory drug in the Egyptian, Indian and Chinese folk medicines, yet its mechanism of action is unclear. Aim of the study To explore its mechanism of action and to correlate it to its biophytochemicals. Materials and methods Various extracts viz. water, 50%, 70%, 80%, and 90% ethanol were prepared from ginger rhizomes. Fractionation of the aqueous extract (AE) was accomplished using Diaion HP-20. In vitro anti-inflammatory activity of the different extracts and isolated compounds was evaluated using protein denaturation inhibition, membrane stabilization, protease inhibition, and anti-lipoxygenase assays. In vivo anti-inflammatory activity of AE was estimated using carrageenan-induced rat paw edema in rats at doses 25, 50, 100 and 200 mg/kg b.wt. Results All the tested extracts showed significant ( p < 0.1) in vitro anti-inflammatory activities. The strongest anti-lipoxygenase activity was observed for AE that was more significant than that of diclofenac (58% and 52%, respectively) at the same concentration (125 μg/ml). Purification of AE led to the isolation of 6-poradol (G1) , 6-shogaol (G2) ; methyl 6- gingerol (G3) , 5-gingerol (G4) , 6-gingerol ( G5) , 8-gingerol (G6) , 10-gingerol (G7) , and 1-dehydro-6-gingerol ( G8 ). G1 , G2 and G8 exhibited potent activity in all the studied assays, while G4 and G5 exhibited moderate activity. In vivo administration of AE ameliorated rat paw edema in a dose-dependent manner. AE (at 200 mg/kg) showed significant reduction in production of PGE2, TNF-α, IL-6, monocyte chemoattractant protein-1 (MCP-1), regulated upon activation, normal T-cell expressed and secreted (RANTES), myeloperoxidase (MPO) activity by 60%, 57%, 60%, 41%, 32% and 67%, respectively. AE at 100 and 200 mg/kg was equipotent to indomethacin in reduction of NO x level and in increasing the total antioxidant capacity (TAC). Histopathological examination revealed very few inflammatory cells infiltration and edema after administration of AE (200 mg/kg) prior to carrageenan. Conclusions Ginger anti-inflammatory activity is mediated by inhibiting macrophage and neutrophils activation as well as negatively affecting monocyte and leukocyte migration. This was evidenced by the dose-dependent decrease in pro-inflammatory cytokines and chemokines and replenishment the total antioxidant capacity. [ABSTRACT FROM AUTHOR]

Phytochemical investigation of the fruits from Trichilia connaroides led to the isolation of four rare 1,2- seco phragmalin-type limonoids ( 1 – 4 ) with C-7/28 δ -lactone ring along with two known triterpenoids ( 5 – 6 ). An oxygen bridge between C-1 and C-2 formed an rare 9- oxa -tricyclo[3.3.2.1 7,10 ]undecane-2-ene moiety in structure of 1 and 2 , and 3 – 4 possessed a bicyclo[5.2.1 4,10 ]decane skeleton. Their structures were established based on extensive spectroscopic methods and their absolute configurations of 1 were confirmed by TDDFT/ECD calculation and comparative analysis. Compounds 5–6 showed moderate inhibitory effects on Hela tumor cell lines with inhibition ratio values of 83.8% and 87.1% at concentration 40 μM, respectively. [ABSTRACT FROM AUTHOR]

Taxifolin is a flavonoid. It has been used as a chemopreventive agent and supplement. It may have some beneficial effects to treat prostate cancer by suppressing androgen production in Leydig cells. The objective of the present study was to study the effects of taxifolin on androgen production of rat Leydig cells isolated from immature testis and some rat and human testosterone biosynthetic enzyme activities. Rat Leydig cells were incubated with 100 μM taxifolin without (basal) or with 10 ng/ml luteinizing hormone (LH), 10 mM 8-bromoadenosine 3′,5′-cyclic monophosphate (8BR), and steroid enzyme substrates (20 μM): 22R-hydroxychloesterol, pregnenolone, progesterone, and androstenedione. The medium concentrations of 5α-androstane-3α, 17β-diol (DIOL) and testosterone were measured. Taxifolin significantly suppressed basal, LH-stimulated, 8BR-stimulated, pregnenolone-mediated, and progesterone-mediated androgen production by Leydig cells. Further study demonstrated that taxifolin inhibited rat 3β-hydroxysteroid dehydrogenase and 17α-hydroxylase/17, 20-lyase with IC 50 values of 14.55 ± 0.013 and 16.75 ± 0.011 μM, respectively. Taxifolin also inhibited these two enzyme activities in human testis with IC 50 value of about 100 μM. Taxifolin was a competitive inhibitor for these two enzymes when steroid substrates were used. In conclusion, taxifolin may have benefits for the treatment of prostate cancer. [ABSTRACT FROM AUTHOR]

Baccharis retusa, a medicinal Brazilian plant from Asteraceae, has been used in Brazilian folk medicine to treatment of several illnesses, including parasitic diseases. Bioactivity-guided fractionation of the n-hexane extract from the aerial parts of B. retusa resulted in the isolation and characterization of three active related diterpenes: ent -15β-senecioyl-oxy-kaur-16-en-19-oic acid ( 1 ), ent -kaur-16-en-19-oic ( 2 ) and ent -16-oxo-17-nor-kauran-19-oic ( 3 ) acids. The structures of isolated compounds were defined by spectroscopic analysis, including NMR and HRESIMS. Antitrypanosomal activity of 1–3 was performed against cell-derived trypomastigotes using the colorimetric resazurin assay. The obtained results demonstrated that isolated compounds displayed a reduced toxicity against NCTC cells and were effective against the trypomastigote forms of T. cruzi with IC 50 values of 3.8 μM ( 1 ), 75.3 μM ( 2 ) and 44.2 μM ( 3 ). Additionally, compound 3 displayed activity against amastigote forms of T. cruzi with IC 50 of 83.2 μM. Compound 1 displayed the highest selectivity index (SI) when considered the trypomastigote forms, and its effect in the plasma membrane of parasite was evaluated using the fluorescent probe SYTOX Green. A considerable permeabilization (57%) in the membrane of the parasite was observed when compared to the untreated trypomastigotes. These data demonstrate, for the first time, the antitrypanosomal activity and mechanism of action of 1 and related compounds 2 and 3 , obtained from aerial parts of B. retusa . [ABSTRACT FROM AUTHOR]

Chemical investigation of Euphorbia helioscopia resulted in the identification of seven new triterpenes named euphorbatrine A-G ( 1 – 7 ), including two (19 α H)-lupane ( 1 – 2 ) and three (9 β H)-lanostane ( 4 – 6 ), which are very rare metabolites from plants. Their structures were elucidated by extensive spectroscopic analysis, X-ray crystallography and chemical methods. Compounds 5 and 6 triggered apoptosis of Hela −/− cell with EC 50 values of 1.59 ± 0.25 and 26.48 ± 0.78 μM, respectively. [ABSTRACT FROM AUTHOR]

Twenty natural stilbenoids ( 1 − 20 ), including seven new stilbenoids ( 2 , 4 – 7 , 19 , 20 ) and thirteen known stilbenoids ( 1 , 3, 8 – 18 ), were isolated from the stem barks of Acanthopanax leucorrhizus , and six modified stilbenoid derivatives ( 1a , 2a , 4a , 4b , 7a and 17a ) were obtained via methylation, demethylation and isopentenylation of the corresponding isolates ( 1 , 2 , 4 , 7 and 17 ). These stilbenoids were structurally characterized by comprehensive analysis of their spectroscopic data and comparison with literature information, and evaluated for their cytotoxic activities against three human tumor cell lines (leukemia HL-60, hepatoma SMMC-7721 and breast carcinoma MCF-7) in vitro by MTT assay. The results showed that compounds 1a , 4a and 4b showed potent selective cytotoxicity against SMMC-7721 ( IC 50 = 10.16 ± 1.95 μM and 9.76 ± 1.32 μM) and MCF-7 ( IC 50 = 10.72 ± 2.78 μM) cell lines. The cytotoxic evaluation of these structurally modified stilbenoid derivatives have led to the establishment of a structure-activity relationship. [ABSTRACT FROM AUTHOR]

Four new naphthoquinones ( 1 – 4 ), named rhinacanthins S ( 1 ), T ( 2 ), U ( 3 ) and V ( 4 ), together with 13 known naphthoquinones were isolated from the leaf extract of Rhinacanthus nasutus . The structures of isolated compounds were elucidated by spectroscopic methods, especially 1D and 2D NMR spectroscopy and mass spectrometry. Rhinacanthin S ( 1 ) exhibited acetylcholinesterase inhibition activity with a % inhibition value of 48.04 ± 3.25. The known rhinacanthin A ( 5 ) showed cytotoxicity against a MCF-7 cell line with an IC 50 value of 8.79 μM, while rhinacanthin N ( 15 ) was active against the NCI-H187 cell line with an IC 50 = 2.24 μM and Vero cells (IC 50 = 3.00 μM) [ABSTRACT FROM AUTHOR]

Resveratrol, a common polyphenol, has extensive pharmacological activities. Resveratrol inhibits some steroid biosynthetic enzymes, indicating that it may block neurosteroid synthesis. The objective of the present study is to investigate the inhibition of resveratrol on neurosteroidogenic enzymes rat 5α-reductase 1 (SRD5A1), 3α-hydroxysteroid dehydrogenase (AKR1C9), and retinol dehydrogenase 2 (RDH2). The IC 50 values of resveratrol on SRD5A1, AKR1C9, and RDH2 were > 100 μM, 0.436 ± 0.070 μM, and 4.889 ± 0.062 μM, respectively. Resveratrol competitively inhibited rat AKR1C9 and RDH2 against steroid substrates. Docking showed that resveratrol bound to the steroid binding pocket of AKR1C9. It exerted a mixed mode on these AKR1C9 and RDH2 against cofactors. In conclusion, resveratrol potently inhibited rat AKR1C9 and RDH2 to regulate local neurosteroid levels. [ABSTRACT FROM AUTHOR]

Four new labdane diterpenoids, 12 β -hydroxy-15-norlabda-8(17),13(14)-dien-16-oic acid ( 1 ), ( E )-15-ethoxy-15-methoxylabda-8(17),12-dien-16-al ( 2 ), ( E )-15 α -ethoxy-14 α -hydroxylabda-8(17),12-dien-16-olide ( 3 ), and 15-ethoxy-12 β -hydroxylabda-8(17),13(14)-dien-16,15-olide ( 4 ) were isolated from the methanol extract of Curcuma amada rhizomes collected in Myanmar, together with 13 known analogs. Their structures were elucidated by extensive spectroscopic techniques. All of the isolates were evaluated for their antiproliferative activities against a small panel of five different human cancer cell lines (A549, human lung cancer; HeLa, human cervical cancer; MCF7, human breast cancer; PANC-1 and PSN-1, human pancreatic cancer). Among the isolates, compounds 2 − 4 , 7 , 8 , 12 , and 17 showed mild antiproliferative activities with IC 50 values ranging from 19.7 to 96.1 μM. ( E )-14-Hydroxy-15-norlabda-8(17),12-dien-16-al ( 11 ) exhibited strong antiproliferative activities selectively against HeLa, PANC-1, and PSN-1 cells, with IC 50 values of 5.88, 1.00, and 3.98 μM, respectively. These potencies were comparable to those of the positive control, 5-fluorouracil. [ABSTRACT FROM AUTHOR]

Five new isocoumarin derivatives, pestalactone A–C ( 1–3 ) and pestapyrone D–E ( 4–5 ), together with two known compounds ( 6–7 ) were isolated from the solid cultures of the endophytic fungus Pestalotiopsis sp. obtained from Photinia frasery . Their structures were mainly determined by extensive spectroscopic analysis, Mo 2 (OCOCH 3 ) 4 -induced electronic circular dichroism (ECD), and ECD calculation. Compounds 1 and 2 were rare isocoumarin derivatives and derived from distinctive polyketide pathways. Compound 3 exhibited potent antifungal activity against Candida glabrata (ATCC 90030) with an MIC 50 value of 3.49 ± 0.21 μg/mL. [ABSTRACT FROM AUTHOR]

Five new limonoids, named sundarbanxylogranins A-E ( 1 – 5 ), were isolated from the seeds of an Indian mangrove, Xylocarpus granatum , collected from mangrove swamps of the Sundarbans, India. The structures of these limonoids were established by HRESIMS and NMR spectroscopic data. Sundarbanxylogranin A ( 1 ) is a rare limonoid containing a bicyclo[5.2.1]dec-3-en-8-one scaffold as the ring A/B-fused core; whereas sundarbanxylogranin B ( 2 ) is a typical mexicanolide with an 8α, 30α-epoxy ring. Sundarbanxylogranins C-E ( 3 – 5 ), among which the former two possess a 29-OMe group with different orientation, belong to a small group of limonoids containing a C 1 - O -C 29 oxygen bridge. Sundarbanxylogranin B exhibited moderate anti-HIV activity with an IC 50 value of 23.14 ± 1.29 μM and a CC 50 value of 78.45 ± 1.69 μM, respectively. [ABSTRACT FROM AUTHOR]

Three propolis samples were collected from different regions of Ecuador (Quito, Guayaquil and Cotacachi) and their methanolic extracts were prepared. Preliminary information supplied by TLC and NMR data, allowed us to define two main types of propolis: Cotacachi propoli sample (CPS), rich in flavonoids and Quito and Guayaquil samples (QPS and GPS) containing triterpenic alcohols and acetyl triterpenes as the main constituents. Two different approaches based on RP-HPLC preparative procedure and NMR structural determination (CPS) and GC–MS analysis (QPS and GPS) were successfully used for the chemical characterization of their major compounds. All three propolis extracts were able to inhibit Leishmania amazonensis growth but propolis sample rich in flavonoids was the most active (IC 50 = 17.1 ± 1.7 μg/mL). In the literature this is the first study on propolis from Ecuador. [ABSTRACT FROM AUTHOR]

A set of PEGylated derivatives of oleanolic and maslinic acids has been semi-synthesised, attaching ethylene glycol, diethylene glycol, triethylene glycol or tetraethylene glycol to the C-28 carboxyl group of these natural triterpenes and some derivatives. Another set of PEGylated derivatives has been semi-synthesised by connecting the same four ethylene glycols to the hydroxyl groups of the A ring of these triterpenic acids, through a carbonate linker, by reaction with trichloromethyl chloroformate. The aqueous solubility of some of these PEGylated derivatives has been compared with that of maslinic acid. The cytotoxic effects of 28 triterpenic PEGylated derivatives in three cancer-cell lines (B16-F10, HT29, and Hep G2) have been assayed. The best results have been achieved with the HT29 cell line, and specifically with the oleanolic acid derivatives having ethylene glycol or tetraethylene glycol attached to the C-28 carboxyl group, which are approximately 27-fold more effective than their natural precursor. Eight PEGylated derivatives have been selected to compare the cytotoxicity results in the HT29 cancer-cell line with those of a non-tumour cell line of the same tissue (IEC-18), four of which were less cytotoxic in the non-tumour cell line. These compounds showed apoptotic effects on treated cells, with percentages of total apoptosis between 20% and 53%, relative to control, at 72 h and IC 50 concentration, and between 29% to 62%, relative to control, for the same time and IC 80 concentration. We have also found that with the treatment of these compounds in HT29 cancer cells, cell-cycle arrest occurred in the G0/G1 phase. Finally, we have also studied changes in mitochondrial membrane potential during apoptosis of HT29 cancer cells, and the results suggest an activation of the extrinsic apoptotic pathway for these compounds. [ABSTRACT FROM AUTHOR]

Three new 2-phenylnaphthalene derivatives, cherrevenaphthalenes A–C ( 1–3 ), and a new polyoxygenated cyclohexene derivative, (−)-uvaribonol F ( 4 ) together with six known compounds, 5–10 , were isolated from the stem and root extracts of Uvaria cherrevensis (Annonaceae). The structures of all isolated compounds were elucidated by spectroscopic analysis. The structures of 3 and 4 were further confirmed by single crystal X-ray diffraction methods. Compound 2 exhibited modest antiplasmodial activity against the P. falciparum stains TM4/8.2 and K1CB1 with IC 50 values of 18.8 ± 3.63 and 23.4 ± 4.08 μM, respectively, and weak cytotoxicity to a Vero cell line. Furthermore, compound 4 displayed cytotoxic activity against a KB cell line with an IC 50 value of 22.1 ± 0.42 μM but was non-cytotoxic to the Vero cell line. Compound 5 revealed stronger cytotoxicity towards the KB cell line, with an IC 50 value of 5.05 ± 0.86 μM and was nearly equally cytotoxic to the Vero cell line. [ABSTRACT FROM AUTHOR]

A series of novel β -elemene isopropanolamine derivatives were synthesized and evaluated for their antitumor activity. The results indicated that all of the compounds showed stronger antiproliferative activities than β -elemene as well as improved aqueous solubility. In particular dimer 6q showed the strongest cytotoxicity against four tumor cell lines (SGC-7901, HeLa, U87 and A549) with IC 50 values ranging from 4.37 to 10.20 μM. Moreover, combination of 6q with cisplatin exhibited a synergistic effect on these cell lines with IC 50 values ranging from 1.21 to 2.94 μM, and reversed the resistance of A549/DPP cells with an IC 50 value of 2.52 μM. The mechanism study revealed that 6q caused cell cycle arrest at the G2 phase and induced apoptosis of SGC-7901 cells through a mitochondrial-dependent apoptotic pathway. Further in vivo study in H22 liver cancer xenograft mouse model validated the antitumor activity of 6q with a tumor inhibitory ratio (TIR) of 60.3%, which was higher than that of β -elemene (TIR, 49.1%) at a dose of 60 mg/kg. Altogether, the potent antitumor activity of 6q in vitro and in vivo warranted further preclinical investigation for potential anticancer chemotherapy. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Leaves from the plant species studied herein are traditionally used in northern Nigeria against various protozoan infections. However, none of these herbal preparations have been standardized, nor have their toxicity to mammalian cells been investigated. In search of improved and non-toxic active antiprotozoal principles that are not cross-resistant with current anti-parasitics, we here report the results of the in vitro screening of extracts from seven selected medicinal plant species ( Centrosema pubescens , Moringa oleifera , Tridax procumbens , Polyalthia longifolia , Newbouldia laevis, Eucalyptus maculate , Jathropha tanjorensis ), used traditionally to treat kinetoplastid infections in Nigeria, and the isolation of their bioactive principles. Aim of the study To investigate the efficacies of medicinal plant extracts, and of compounds isolated therefrom, against kinetoplastid parasites, assess cross-resistance to existing chemotherapy, and assay their toxicity against mammalian cells in vitro. Material and methods Plants were extracted with hexane, ethyl acetate and methanol. Active principles were isolated by bioassay-led fractionation, testing for trypanocidal activity, and identified using NMR and mass spectrometry. EC 50 values for their activity against wild-type and multi-drug resistant Trypanosoma brucei were obtained using the viability indicator dye resazurin. Results Seven medicinal plants were evaluated for activity against selected kinetoplastid parasites. The result shows that crude extracts and isolated active compounds from Polyalthia longifolia and Eucalyptus maculata , in particular, display promising activity against drug-sensitive and multi-drug resistant Trypanosoma brucei . The EC 50 value of a clerodane (16α-hydroxy-cleroda-3,13(14)-Z-dien-15,16-olide) isolated from Polyalthia longifolia was as low as 0.38 µg/mL, while a triterpenoid (3β,13β-dihydroxy-urs-11-en-28-oic acid) isolated from Eucalyptus maculata displayed an EC 50 of 1.58 µg/mL. None of the isolated compounds displayed toxicity towards Human Embryonic Kidney cells at concentrations up to 400 µg/mL. In addition, the isolated compounds were active against Leishmania mexicana , as well as against T. congolense . Conclusion We have isolated a clerodane compound from Polyalthia longifolia that shows low toxicity, no cross-resistance with current treatments, and promising activity against both human-infective and veterinary Trypanosoma species. [ABSTRACT FROM AUTHOR]

Ethnopharmacological importance The hemiparasitic species Psittacanthus plagiophyllus Eichl. (Loranthaceae), also known as erva de passarinho , is used in folk medicine in the Santarém region in the state of Pará, Brazil, to treat gastritis and a variety of inflammatory disorders. In view of the lack of pharmacological studies on this species in the literature and the fact that it is used constantly by the local population, this study sought to standardize the extract of the leaves of P. plagiophyllus (AEPp) and to assess its anti-inflammatory potential in in vivo tests. Material and methods Quality control and standardization of AEPp were performed following the 5th edition of the Brazilian Pharmacopeia. To assess the in vivo anti-inflammatory activity of AEPp, the carrageenan-induced and dextran-induced rat paw edema models were initially used. To investigate the effect of AEPp on carrageenan-induced leukocyte recruitment and exudate production, the air pouch inflammation model was used. In addition, the antioxidant activity of AEPp was assessed in vitro by the DPPH radical scavenging method. Results The chromatographic profile of AEPp indicated the presence of flavonoids, coumarins and hydrolyzable and condensed tannins. Measurement of phenolics revealed the following percentages in the extract: 12.62±0.18% total phenolics, 5.39±0.01% total tannins, 12.54±0.24% hydrolyzable tannins, 8.37±0.32% condensed tannins and 1.23±0.02% total flavonoids. In 500 and 1000 mg/kg doses (p.o.) AEPp had significant edema-inhibiting activity ( p <0.01) in both the models used, suggesting that the extract may act in vascular and cell events in the inflammatory response and exert an inhibitory effect on mediators responsible for edema. In all the doses tested [250, 500 and 1000 mg/kg (p.o.)], AEPp inhibited total leukocyte and neutrophil migration and reduced the amount of exudate in the air pouch in a dose-dependent manner. Both effects were statistically significant ( p <0.01). The extract also reduced the DPPH radical (maximum reduction 93.13±1.71% at a concentration of 75 µg/mL), indicating that it has antioxidant activity. AEPp, therefore, exhibited considerable in vivo anti-inflammatory activity and in vitro antioxidant activity. This may be due to its high phenolics content. Conclusion These findings provide evidence to support the use of P. plagiophyllus in folk medicine to treat inflammatory disorders. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Terminalia bellerica Roxb. fruits are rich in a variety of biologically active ingredients. Tharu and Buksa tribes of Udham Singh Nagar, Uttarakhand, India use grilled fruits of Terminalia bellerica as an effective cure for diarrhea Aim of the study We validated the ethnobotanical claim by comparing the antidiarrheal effect of grilled fruits (GF) with dried fruits (DF). Materials and methods The 50% ethanolic extracts of GF and DF were successively fractionated; the antioxidant and bacterial inhibition activity were studied using DPPH free radical scavenging, anti-lipid peroxidation and broth dilution method respectively. Difference in metabolites of ethyl acetate fractions of GF and DF was analyzed using GC-MS, gallic acid content was determined through HPTLC. Further the in-vivo antidiarrheal effect of ethyl acetate fractions of DF and GF was studied on castor oil induced diarrhea model. Results The ethyl acetate fractions showed potential DPPH free radical scavenging (IC 50 11.13 µg/ml in DF and 8.56 µg/ml in GF), anti-lipid peroxidation and antibacterial activity. The non-targeted metabolic profiling showed higher content of tartaric acid, valeric acid, gallic acid, succinic acid, oxalic acid, malonic acid, malic acid, 1,2,3 trisbenzene, uridine and 11-eicosenoic acid in GF. The HPTLC results indicated that gallic acid content was 2.8 (±0.14) and 4.92 (±0.28) mg/g while ellagic acid content was 4.7 (±0.32) and 4.45 (±0.45) mg/g dry powder in DF and GF respectively. According to i n vivo antidiarrheal activity DF and GF (100 mg/kg oral) inhibited diarrhea by 41.87% and 71.72% respectively. Conclusion Grilling significantly altered the levels of metabolites in T. bellerica fruits which could be responsible for its increased therapeutic potential. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Psidium guajava and Tagetes erecta have been used traditionally to treat gastrointestinal parasites, but their active metabolites and mechanisms of action remain largely unknown. Aim of the study To evaluate the anthelmintic potential of Psidium guajava and Tagetes erecta extracts on Levamisole-sensitive and Levamisole-resistant strains of the model nematode Caenorhabditis elegans. Materials and methods Aqueous extracts of Psidium guajava (PGE) and Tagetes erecta (TEE) were assayed on locomotion and egg-laying behaviors of the wild-type (N2) and Levamisole-resistant (CB193) strains of Caenorhabditis elegans . Results Both extracts paralyzed wild-type and Levamisole-resistant nematodes in a dose-dependent manner. In wild-type worms, TEE 25 mg/mL induced a 75% paralysis after 8 h of treatment and PGE 25 mg/mL induced a 100% paralysis after 4 h of treatment. PGE exerted a similar paralyzing effect on N2 wild-type and CB193 Levamisole-resistant worms, while TEE only partially paralyzed CB193 worms. TEE 25 mg/mL decreased N2 egg-laying by 65% with respect to the untreated control, while PGE did it by 40%. Conclusions Psidium guajava leaves and Tagetes erecta flower-heads possess hydrosoluble compounds that block the motility of Caenorhabditis elegans by a mechanism different to that of the anthelmintic drug Levamisole. Effects are also observable on oviposition, which was diminished in the wild-type worms. The strong anthelmintic effects in crude extracts of these plants warrants future work to identify their active compounds and to elucidate their molecular mechanisms of action. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Campomanesia velutina leaves and branches infusions are used in Brazilian folk medicine to treat diarrhea and to ameliorate intestinal cramps, respectively. Aim of the study Carry out the acute and sub chronic pre-clinical evaluation and thus assess the safety and toxicological potential of the specie. Materials and methods In vivo toxicity was evaluated by acute and sub chronic toxicity assays conducted according to the guidelines of the Brazilian Agency of National Health Surveillance (Agência Nacional de Vigilância Sanitária – ANVISA). For acute toxicity evaluation, a single dose of aqueous extracts from the leaves (AEL) and branches (AEB) of Campomanesia velutina were orally administered to mice at doses of 300, 600 and 1200 mg/kg. Then, the animals were observed for 14 days. In the sub chronic study, the extracts were orally administered to mice for 14 days at doses of 300, 600 and 1200 mg/kg. To assess the toxicological effects, animals were closely observed on general behavior, clinical signs of toxicity, body weight, food and water intake. At the end of the experiment, it was performed biochemical and hematological evaluations, as well as histopathological analysis from the following organs: brain, heart, lungs, liver, stomach, small intestine (section) and left kidney. Preliminary phytochemical analysis was performed using thin layer chromatography (TLC) and colorimetric pharmacognostic tests. Results In oral acute assay, treatment with AEB at the major dose (1200 mg/kg) caused diarrhea, abdominal cramps and tremors in females. These effects were reversed at 4th hour. Normochromic normocytic anemia was observed in males treated with AEL 300 mg/kg and AEB 600 and 1200 mg/kg as well as in females treated with AEB 300 and 1200 mg/kg. The kidney of all treated animals showed moderate inflammation and a few hemorrhagic points. In sub chronic assay, treatment with AEL 600 mg/kg, AEL 1200 mg/kg and AEB 1200 mg/kg caused hyper excitability in females that was not reversed. Treatments also had impact on weight gain and the relative weight of males’ brain was increased on group treated with AEL 300 mg/kg, AEB 300 and AEB 1200 mg/kg. Although changes in hematological parameters were not observed, serum creatinine levels were significantly higher in males treated with AEB 300 mg/kg. Besides, the heart of all treated animals showed intense hyperemia. Preliminary phytochemical analysis revealed the presence of flavonoids, tannins and phenolic compounds. Conclusions Toxicity signs were mainly observed after treatment with AEL and AEB at the two highest tested doses (600 and 1200 mg/kg), suggesting that the extracts are relatively safe at its effective dose (300 mg/kg). However, alterations on hematological and biochemical parameters and on the kidney and heart of the animals were not closely related with the dose, implying caution on its use. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Corn silk or the stigma of Zea mays L. has traditionally been used in weight loss stimulation and treatment of cystitis, urinary infections and obesity. Purple corn silk, rich of polyphenolic substances, was reported on anti-diabetic and anti-obesity effect in animal studies. However, scientific evidence on mechanisms and targets of action of purple corn silk related to adipocyte life cycle has been limited. Aim of study To determine phytochemical compositions and investigate anti-obesity potential of the purple corn silk focusing on interruption of adipocyte life cycle; effect on pre-adipocyte proliferation, adipogenesis, adipocyte lipolysis, and apoptosis. Materials and methods The ethanolic purple corn silk extract (PCS) was prepared and investigated for phytochemical compositions by LC/MS/MS technique and anti-obesity potential using murine 3T3-L1 cell line. Using methyl thiazole tetrazolium (MTT) assay, the effects on pre-adipocytes and adipocyte viability and on pre-adipocytes proliferation at 24-, 48-, and 72-h incubation period were evaluated. In addition, anti-adipogenesis via inhibition on adipocyte differentiation and reduction of total lipid accumulation was evaluated using Oil Red O staining and spectrophotometric methods, respectively. The lipolysis effect was determined by measurement of glycerol released content using glycerol test kit after 48-h treatment of PCS to adipocytes. Apoptosis inductive effect was done by using 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) staining method. Results The polyphenols including anthocyanins, quercetin and phenolic acids and derivatives were found as the major chemical compositions of the PCS. With multiple-stages interruption on the adipocyte life cycle, anti-obesity effect of PCS was interestingly demonstrated. When compared to the control, the PCS at concentration range between 250–1000 μg/mL showed anti-adipogenesis effect as expressing of significant inhibition on pre-adipocyte proliferation at all incubation period (43.52±5.28 – 75.51±9.09%) and significant decreasing of total lipid accumulation at concentration of 500 μg/mL (80.22±6.58%) and 1000 μg/mL (69.62±5.42%). Moreover, the PCS exhibited lipolysis and apoptosis inductive effect with dose dependent manner and significance at concentration of 1000 μg/mL by increase of released glycerol content (173.88±6.13% of the control) and of nuclei condensing and apoptotic bodies (with relative apoptosis induction as 131.74±1.64% of the control). Conclusion Our data has evidenced the anti-obesity potential of PCS related interruption at multiple stages of adipocyte life cycle. Its potency was attributed to inhibition on adipocyte proliferation and adipogenesis as well as induction on lipolysis and apoptosis at high concentration. However, further in vivo investigation should be considered to insist the possibility in applications of PCS in prevention and treatment of obesity. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance The genus Hyptis comprehends almost 400 species widespread in tropical and temperate regions of America. The use of Hyptis spicigera Lam. (Lamiaceae) is reported in traditional medicine due to its gastroprotective, anti-inflammatory and analgesic properties. Aim of the study The rationale of this study was to investigate the potential use of the essential oil of H. spicigera (EOHs) as analgesic. Material and methods The antinociceptive effect of EOHs was verified analyzing acute nocifensive behavior of mice induced by chemical noxious stimuli [i.e., formalin and transient receptor potential (TRP) channels agonists]. We also verified the effects of EOHs on locomotor activity and motor performance in mice. Finally, we investigate the involvement of central afferent C-fibers with EOHs analgesic effect. Results EOHs presented antinociceptive effect at 300 and 1000 mg/kg on formalin-induced pain behavior model, presenting 50% and 72% of inhibition during the first phase (ED 50 =292 mg/kg), and 85% and 100% during de second phase (ED 50 =205 mg/kg), respectively. Temperature of the hind paw was reduced by EOHs treatment in a dose-dependent manner; oedema was diminished only by EOHs 1000 mg/kg. EOHs does not impaired locomotor activity or motor performance. For mice injected with capsaicin, a TRPV1 activator, EOHs (1000 mg/kg, ED 50 =660 mg/kg) showed decreased (63%) nociceptive behavior. When injected with cinnamaldehyde (TRPA1 activator), mice treated with EOHs showed 23%, 43% and 66% inhibition on nociceptive behavior (100, 300 and 1000 mg/kg, respectively; ED 50 402 mg/kg). When mice were injected with menthol (TRPM8 activator), EOHs showed 29%, 59% and 98% inhibition of nociceptive behavior (100, 300 and 1000 mg/kg, respectively; with ED 50 =198 mg/kg. Finally, when desensitized mice were injected with menthol, EOHs (300 mg/kg) does not show antinociceptive effect. Conclusions This study demonstrated the efficacy of EOHs on experimental models of nociception. We have found the involvement of TRP channels V1, A1 and M8 with EOHs activity, which was remarkably potent and efficient in inhibiting pain evoked by menthol, a TRPM8 channel activator. TRPM8 channels from TRPV1+ C-fibers, but not TRPM8+ C-fibers nor TRPM8+ Aδ mechanosensory fibers, mediate EOHs analgesic effects. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Spatholobi Caulis is a traditional blood-activating and stasis-dispelling herb medicine, which has been used to treat diseases related to blood stasis syndrome (BSS) by inhibiting platelet aggregation, stimulate hematopoiesis, etc. It has been demonstrated that pro-angiogenesis could improve BSS. However, the pro-angiogenic activity of Spatholobi Caulis was not well elucidated Aim of study To determine the potential pro-angiogenic activity of Spatholobi Caulis and elucidate its underlying mechanism. The active fractions of Spatholobi Caulis were further screened. Material and methods Gelatin precipitation and reversed-phase liquid chromatography (RPLC) were used to purify the methanol extracts of Spatholobi Caulis , respectively. The RPLC was also used to prepare fractions. Total flavonoids of purified methanol extracts of Spatholobi Caulis (PSC) were determined using ultraviolet spectrophotometry. The morphological observation of subintestinal vessel plexus (SIVs) and tyrosine kinase inhibitor II (VRI)-induced intersegmental blood vessels (ISVs) loss in transgenic zebrafish Tg(fli-1a: EGFP)y1 were selected to evaluate the pro-angiogenic activity of PSC in vivo. Cell proliferation by MTT assay and cell migration assay were used to evaluate the pro-angiogenesis effect of PSC in human umbilical vein endothelial cells (HUVECs) in vitro . Both zebrafish and HUVECs were used in screening active fractions of PSC. The mechanism of PSC promoting angiogenesis were studied by real-time PCR in zebrafish and western blotting in HUVECs. Results Co-treatment PSC dramatically rescued VRI-induced ISVs loss in zebrafish embryos in a dose-dependent manner and 80% of the defective vascular recovered at the concentration of 30 μg/ml compared with VRI-only group. PSC also concentration-dependently increased average sprouting number and diameter of SIVs in zebrafish embryo. Real-time PCR assay proved that PSC significantly restored the down regulation of VEGFRs including Flt-1, Kdr and Kdrl induced by VRI in zebrafish (P<0.001). Furthermore, PSC not only promoted proliferation and migration of normal HUVECs but also ameliorated VRI-induced HUVECs cytotoxicity. Western blotting assay showed that co-treatment of PSC increased the expression of VEGFRs and phosphorylation of MAPKs which decreased by VRI treatment. In addition, quality evaluation experiments showed that the content of total flavonoids of PSC reached 56.36% and the main pro-angiogenic fractions of PSC were F3, F4 and F5 both in zebrafish and HUVECs. Conclusions Our data demonstrated that PSC presented pro-angiogenic activity both in zebrafish and HUVECs, and principal pro-angiogenic active components were likely flavonoids. Thus, the current study provided evidence for the clinical usage of Spatholobi Caulis in promoting blood circulation and removing stasis in traditional Chinese medicine (TCM). [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance The roots, stem and leaves of Morinda lucida are used in some African countries as treatment against different types of fevers including yellow fever, malaria, trypanosomiasis and feverish conditions during child birth. Aim of the study To determine the in vitro cell toxicity and anti-malarial activity of the extracts of stem bark of M. lucida and to identify the secondary metabolites in the extract that may be responsible for this activity. Materials and methods The cell toxicity studies of crude extract [dichloromethane (DCM): Methanol (MeOH) in a ratio of1:1 (v/v)] as well as compounds isolated from the same extract were carried out using human cervix adenocarcinoma cells (HeLa cells); while the anti-malarial activities of the same samples were performed against Plasmodium falciparum strain 3D7 using the parasite lactate dehydrogenase (pLDH) assay. The isolation of the active compounds was carried out using chromatographic techniques (column and thin layer chromatography) where as mass spectrometry (MS), Fourier transform infrared spectroscopy (FTIR) as well as 1D- and 2D- nuclear magnetic resonance (NMR) analyses were employed in the characterisation and identification of the isolated secondary metabolites. Results The pLDH and cell toxicity assays for the crude extract and the fractions of M. lucida indicated that some fractions reduced the malaria parasite viability by approximately 50% at 100 μg/mL and they were not significantly cytotoxic. An IC 50 done on the crude extract gave a value of 25 μg/mL. The % cell viability for the crude extract in cell toxicity assay remained at 100%. Seven chemical constituents i.e. asperuloside ( 1 ), asperulosidic acid ( 2 ), stigmasterol ( 3a ), β-sitosterol ( 3b ), cycloartenol ( 3c ), campesterol ( 3d ) and 5,15-O-dimethylmorindol ( 4 ) were isolated from the DCM-MeOH extract of stem bark. The isolated compounds tested were not that active by themselves individually at 20 μM but their activities were increased when the isolated compounds were combined. As seen when compounds 2, 3 and 4 (% viability: 93, 123 and 101 respectively) were combined yielding an IC 50 value of 17 μM. Furthermore, this is the first report of compounds 1, 2, 3c, 3d and 4 isolated from M. lucida . Conclusion The crude extract completely suppressed the growth of P. falciparum . This indicates that the crude extract contains many compounds that might be acting in synergy. The observed activity of the crude extract and the samples containing a mixture of different compounds support the traditional use of M. lucida for the treatment of malaria. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance The root extract of Albizia zygia (DC.) J.F. Macbr. (Leguminosae-Mimosoideae) is traditionally used in the management of pain and fever. However, little scientific data exists in literature to support its use. Aim of study The present study evaluated the anti-nociceptive and antipyretic properties of the hydroethanolic extract of the roots of Albizia zygia in animal models. Materials and methods The analgesic effects were investigated in chemical (acetic acid-induced abdominal writhing and formalin tests), thermal (tail-immersion test) and mechanical (carrageenan-induced hyperalgesia) pain models. Possible mechanisms of anti-nociception were also assessed with antagonists in the formalin test. The anti-pyretic effect was evaluated using the baker yeast-induced pyrexia model in young rats. Results The extract (30–300 mg/kg, p.o .) and positive controls, diclofenac (3–30 mg/kg, i.p.) and morphine (1–10 mg/kg, i.p.), significantly (at least P <0.01) attenuated acetic acid-induced visceral pain, formalin- induced paw pain (both neurogenic and inflammatory), thermal pain as well as carrageenan-induced mechanical hyperalgesia in animals. The anti-nociceptive effect of the extract was reversed (at least P <0.05) by the pre-emptive administration of naloxone and atropine; the administration of theophylline, however, exhibited no significant ( P >0.05) inhibition of anti-nociception. The extract (30–300 mg/kg, p.o ) and paracetamol (15–150 mg/kg, p.o. ) both reversed yeast-induced pyrexia in rats with ED 50 values of 48.59±2.59 and 26.19±1.33 mg/kg respectively. Conclusion The findings indicate that the extract possesses significant anti-nociceptive and antipyretic effects which justify its traditional use in the management of pain and fever. Also, anti-nociceptive effect of the extract involves opioidergic and muscarinic cholinergic mechanisms. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance The ripe seeds of Herpetospermum caudigerum have been used in Tibetan folk medicine for treatment of bile or liver diseases including jaundice, hepatitis, intumescences or inflammation. Previously reports suggested that the seed oil and some lignans from H. caudigerum exhibited protective effects against carbon tetrachloride (CCl 4 )-induced hepatic damage in rats, which may be related to their free radical scavenging effect. However, the protective effect of H. caudigerum against cholestasis is still not revealed. The aim of the present study was to investigate the pharmacological effect and the chemical constituents of the petroleum ether extract (PEE) derived from H. caudigerum against α-naphthylisothiocyanate (ANIT)-induced acute cholestasis in rats. Materials and methods Male cholestatic Sprague-Dawley (SD) rats induced by ANIT (60 mg/kg) were orally administered with PEE (350, 700 and 1400 mg/kg). Levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-Glutamyl transpeptidase (γ-GTP), total bilirubin (TBIL), direct bilirubin (DBIL) and total bile acid (TBA), as well as bile flow, and histopathological assay were evaluated. Hepatic malondialdehyde (MDA), myeloperoxidase (MPO), superoxide dismutase (SOD), glutathione S-transferase (GST), and nitric monoxide (NO) in liver were measured to explore the possible protective mechanisms. Phytochemical analysis of PEE was performed by gas chromatography-mass spectrometer (GC-MS). Results PEE have exhibited significant and dose-dependent protective effect on ANIT-induced liver injury by reduce the increases in serum levels of ALT, AST, ALP, γ-GTP, TBIL, DBIL and TBA, restore the bile flow in cholestatic rats, and reduce the severity of the pathological tissue damage induced by ANIT. Hepatic MDA, MPO and NO contents in liver tissue were reduced, while SOD and GST activities were elevated in liver tissue. 49 compounds were detected and 39 of them were identified by GC-MS analysis, in which long-chain fatty acids were the main constituents. Conclusions PEE exhibited a dose-dependently protective effect on ANIT-induced liver injury in cholestatic rats with the potential mechanism of attenuated oxidative stress in the liver tissue, and the possible active compounds were long-chain fatty acids. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance World Health Organization (WHO) has recognized the contribution of traditional medicine in the primary health care system. WHO also considers neurological disorders as one of the prime concern to the public health. Inadequate and inaccessible health facilities in the rural and underprivileged areas have forced the various tribal and non-tribal communities to use ethno-phyto and ethno-zoo therapeutics against different ailments. Chota Nagpur Plateau, India is one of the richest in India in terms of plant and animal diversity, and houses a number of indigenous communities often practising their own ethnobiological medicines. These undocumented indigenous treasure must be preserved and could be utilized in discovering lead compounds against human ailments including neuroprotective and psychoactive disorders. Materials and methods A number of ethnobiological surveys were carried out during 2012–2015 in the three districts of the Bengal part of Manbhum region, Chota Nagpur Plateau via conducting open and semi-structured interviews among the traditional medicine men (65:59 males and 6 females aged between 22 and 85 years). This dataset was statistically analysed by calculating seven quantitative indexes such as use value (UV), informants' consensus factor (ICF), fidelity level (FL), relative importance (RI), cultural importance index (CII), index of agreement on remedies (IAR) and cultural agreement index (CAI). Results Sixty five traditional medicine men (TMM) reported 47 traditional formulations involving 58 plant species and 17 animal species used against 13 neurological and psychological disorders and as pro-cognitives. Fabaceae was represented by the highest number of medicinal plant species (9 species) whereas herbs (48%) and roots (28%) represented the most common plant habit and plant part, respectively. Quantitative ethnobiology revealed the frequency and coherence of ethnomedicinal reports. The plant Piper nigrum (1.35) and the animal, Spilostethus hospes (0.50) showed the highest UV. Among the ethnobotanicals, maximum RI value was noted for Hemidesmus indicus (0.91) and IAR was found to be maximum (1.00) for 8 plant species. Among the ethnozoologicals, Spilostethus hospes (0.67) displayed maximum RI value whereas Axis axis (2.32) displayed maximum CAI value. Conclusion TMM possessed knowledge on preparations, dose dependence, administration, safety, possible toxicity and side effects of using traditional formulations. TMM practising age old traditional therapeutics serve as alternative medical option for the underprivileged rural ethnic people in these remote areas of the country. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Huperzia squarrosa (Forst.) Trevis is used in traditional medicine for improving memory deficits. Alkaloids, triterpenoids, flavonoids are main bioactive compounds of Huperzia squarrosa (Forst.) Trevis. Aim of the study This study aimed to investigate the antioxidant, AChE inhibitory activities in vitro of differents fraction of Huperzia squarrosa (Forst.) Trevis extract and neuroprotective effects of EtOAc fraction on scopolamine-induced cognitive impairment in mice. Materials and methods Antioxidant activity was measured by DPPH assay. AChE inhibitory effect in vitro and detail kinetic inhibition mechanism was evaluated by Ellman's assay. For in vivo assay, mice were administrated orally EtOAc fraction (150 and 300 mg/kg) for fourteen days, and injected scopolamine at a dose of 1 mg/kg intraperitoneally for four days to induce memory injured. The memory behaviors were evaluated using the Morris water maze. ACh levels were measured in brain tissue. Superoxide dismutase (SOD), glutathione peroxidase (GPx) activities, malondialdehyde and protein thiol groups were also evaluated in the brains. Results Our data also demonstrated that EtOAc fraction had the strongest antioxidant with an IC 50 value of 9.35±1.68 µg/mL and AChE inhibitory activity with an IC 50 value of 23.44±3.14 μg/mL in a concentration-dependent manner. Kinetic inhibition analysis indicated that EtOAc fraction was mixed inhibition type with Ki (representing the affinity of the enzyme and inhibitor) was 34.75±1.42 µg/mL. Scopolamine significantly increased the escape latency time, reduced the crossings number, and swimming time in the target quadrant, while EtOAc fraction reversed these scopolamine-induced effects. EtOAc fraction significantly increased levels of acetylcholine in the brain. EtOAc fraction also significantly decreased oxidative stress in mice. Conclusion Our data suggest that EtOAc fraction of Huperzia squarrosa extract exhibited a strong neuroprotective effect on cognitive impairment, and may be a potential candidate for the treatment of Alzheimer. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Cancer has proceeded to surpass one of the most chronic illnesses to be the major cause of mortality in both the developing and developed world. Garcinia mangostana L. (mangosteen, family Guttiferae) known as the queen of fruits, is one of the most popular tropical fruits. It is cultivated in Southeast Asian countries: Malaysia, Indonesia, Sri Lanka, Burma, Thailand, and Philippines. Traditionally, numerous parts of G. mangostana have been utilized to treat various ailments such as abdominal pain, haemorrhoids, food allergies, arthritis, leucorrhoea, gonorrhea, diarrhea, dysentery, wound infection, suppuration, and chronic ulcer. Aim of study Although anticancer activity has been reported for the plant, the goal of the study was designed to isolate and characterize the active metabolites from G. mangostana and measure their cytotoxic properties. In this research, the mechanism of antiproliferative/cytotoxic effects of the tested compounds was investigated. Materials and methods The CHCl 3 fraction of the air-dried fruit hulls was repeatedly chromatographed on SiO 2 , RP 18 , Diaion HP-20, and polyamide columns to furnish fourteen compounds. The structures of these metabolites were proven by UV, IR, 1D, and 2D NMR measurements and HRESIMS. Additionally, the cytotoxic potential of all compounds was assessed against MCF-7, HCT-116, and HepG2 cell lines using SRB-U assay. Antiproliferative and cell cycle interference effects of potentially potent compounds were tested using DNA content flow cytometry. The mechanism of cell death induction was also studied using annexin-V/PI differential staining coupled with flow cytometry. Results The CHCl 3 soluble fraction afforded two new xanthones: mangostanaxanthones V ( 1 ) and VI ( 2 ), along with twelve known compounds: mangostanaxanthone IV ( 3 ), β -mangostin ( 4 ), garcinone E ( 5 ), α -mangostin ( 6 ), nor-mangostin ( 7 ), garcimangosone D ( 8 ), aromadendrin-8-C- β -D-glucopyranoside ( 9 ), 1,2,4,5-tetrahydroxybenzene ( 10 ), 2,4,3`-trihydroxybenzophenone-6-O- β -glucopyranoside ( 11 ), maclurin-6-O- β -D-glucopyranoside (rhodanthenone) ( 12 ), epicatechin ( 13 ), and 2,4,6,3`,5`-pentahydroxybenzophenone ( 14 ). Only compound 5 showed considerable antiproliferative/cytotoxic effects with IC 50 's ranging from 15.8 to 16.7 µM. Compounds 3 , 4 , and 6 showed moderate to weak cytotoxic effects (IC 50 's ranged from 45.7 to 116.4 µM). Using DNA content flow cytometry, it was found that only 5 induced significant cell cycle arrest at G 0 /G 1 -phase which is indicative of its antiproliferative properties. Additionally, by using annexin V-FITC/PI differential staining, 5 induced cells killing effect via the induction of apoptosis and necrosis in both HepG 2 and HCT116 cells. Compound 3 produce necrosis and apoptosis only in HCT116 cells. On contrary, 6 induced apoptosis and necrosis in HepG 2 cells and moderate necrosis in HCT116 cells. Conclusion Fourteen compounds were isolated from chloroform fraction of G. mangostana fruit hulls. Cytotoxic properties exhibited by the isolated xanthones from G. mangostana reinforce the avail of it as a natural cytotoxic agent against various cancers. These evidences could provide relevant bases for the scientific rationale of using G. mangostana in anti-cancer treatment. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Following Venezuelan traditional medicine, females with heavy menstrual blood loss (menorrhagia) drink Brownea grandiceps Jacq. flowers ( BG ) decoctions to reduce the bleeding. In a previous study, we demonstrated that BG aqueous extract ( E ) possesses a potent anti-fibrinolytic activity capable of inhibiting plasmin, the main serine-protease that degrades fibrin. It is widely known that plasmin inhibitors are often used as anti-fibrinolytics to reduce bleeding during surgeries with high risk of blood loss such as cardiac, liver, vascular, tooth extraction and large orthopedic procedures, as well as for menorrhagia treatments. The aim of this work was to isolate and characterize from BGE the compound responsible for the reported anti-fibrinolytic activity. Materials and methods A decoction of BG was prepared; then it was homogenized, centrifuged and lyophilized to obtain BGE. Subsequently the extract was fractionated by gel filtration and reverse phase using HPLC and the active compound was characterized by MALDI-ToF MS. The kinetic parameters of anti-plasmin activity were evaluated by an amidolytic assay using a chromogenic substrate; also the anti-plasmin activity was estimated by fibrin plate method. Data were analyzed by nonparametric statistics. Results The active compound was a condensed tannin denominated Browplasminin, which is capable of inhibiting the plasmin activity in a dose-dependent manner when measured in fibrin plates or by the amidolytic activity method; it also has a minor effect on the FXa activity. However, it does not affect the activity of other serine-proteases such as trypsin, t-PA or u-PA. Browplasminin consists predominately of heteroflavan-3-ols of catechin with B-type linkages, and extents up to heptadecamers (~ 5000 Da), with hexose residues attached to the polymer that presents a high degree of galloylation. Its IC 50 for plasmin was 47.80 μg/mL and for FXa was 237.08 μg/mL, while the Ki were 0.76 and 61.61 μg/mL for plasmin and FXa, respectively. Conclusions The overall outcome of this study suggests that Browplasminin could be responsible for reducing heavy menstrual bleeding in women because its kinetic parameters showed that is a good plasmin inhibitor. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Licochalcone A (LCA) is a characteristic chalcone that is found in licorice, which is a traditional medicinal plant. In traditional medicine, LCA possesses many potential biological activities, including anti-parasitic, anti-inflammatory and antitumor activities. Aim of the study To determine the antioxidant activity of LCA and, on this basis, to investigate the role of its anticancer activity. Materials and methods To validate the antioxidant activity of LCA, the proteins SOD, CAT and GPx1 were analyzed using western blotting and cellular antioxidant activity (CAA) assays. Oxidative free radicals are associated with cancer cells. Therefore, the anticancer activity of LCA was also evaluated. To assess the anticancer activity, cell viability assays were performed and apoptosis was evaluated. In addition, MAPK-related proteins were analyzed using western blotting. Results The experimental data showed that the EC 50 of LCA is 58.79±0.05 μg/mL and 46.29±0.05 μg/mL under the two conditions tested, with or without PBS. In addition, LCA at a concentration of approximately 2–8 μg/mL can induce the expression of SOD, CAT and GPx1 proteins. Further, LCA inhibits the growth of HepG2 cells through cell proliferation arrest and the subsequent induction of apoptosis, and LCA attenuated the p38/JNK/ERK signaling pathway in a dose-dependent manner. Conclusion The results showed that LCA suppresses the oxidation of cells and markedly inhibits the proliferation of cancer cells. These findings confirm the traditional use of LCA in folk medicine. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance As an effort to identify newer anti-obesity lead(s) we have selected 13 plant materials from the six plant species which have been reported in Indian Ayurvedic medicine as remedy against complications affecting glucose and lipid homeostasis. Aim of the study In vitro screening of six Indian Ayurvedic medicinal plants on anti-adipogenic and pancreatic lipase (PL) inhibition potential followed by bioactivity guided isolation from most active plant material. Materials and methods In vitro anti-adipogenic assay using 3T3-L1 preadipocytes and pancreatic lipase (PL) inhibition assay were performed for hexanes, dichloromethane, ethyl acetate and methanolic extracts of all the plant materials. Bioactivity guided isolation approach was used to identify active constituent for anti-adipogenesis and PL inhibition assay. Inhibition of lipid accumulation and adipogenic transcription factor was measured by oil Red ‘O’ staining and quantitative real-time PCR method respectively. Results Ethyl acetate extract of Oroxylum indicum bark was found to be most active in screening of anti-adipogenesis (59.12±1.66% lipid accumulation as compared to control at 50 μg/mL dose) and PL inhibition (89.12±6.87% PL inhibition at 250 μg/mL dose) assays. Further, three bioactive flavonoids were isolated and identified as oroxylin A, chrysin and baicalein from O. indicum bark. Oroxylin A, chrysin, and baicalein were inhibited lipid accumulation in 3T3-L1 preadipocytes (75.00±5.76%, 70.21±4.23% and 77.21±5.49% lipid accumulation respectively in comparison to control at 50 μM dose) and PL enzyme (69.86±2.96%, 52.08±2.14% and 45.06±2.42% PL inhibition respectively at 250 μg/mL dose). In addition, oroxylin A and chrysin also inhibited PPARγ and C/EBPα, major adipogenic transcription factors, in 3T3L-1 preadipocytes during adipogenesis process at 50 μM dose. Conclusion The present study augurs the anti-obesity potential of well practiced Ayurvedic herb O. indicum and its flavonoids. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Arishtas are Ayurvedic formulation made with decoction of herbs. Arjunarishta formulation is being used in Ayurveda for cardio-protective activity. Ashwagandharishta formulation possesses antioxidant, anti-atherosclerotic and anti-stress properties. Ridayarishta, a novel empirical formulation was prepared using combination of selected ingredients from these two formulations to support healthy heart functions and to reduce stress. Aim of the Study Aim of the Study was to investigate herb-drug interaction (HDI) of Ridayarishta formulation through human hepatic cytochrome P450 (CYP450) enzyme inhibition assay. Materials and methods Ridayarishta formulation was phyto-chemically standardized against arjunolic acid, arjunetin, berberine, piperine, resveratrol and withaferin-A using high performance thin layer chromatography (HPTLC) analysis. The formulation was standardized with respect to ethanol by gas chromatographic (GC) analysis. HDI was evaluated with Ridayarishta formulation and amlodipine besilate, atenolol, atorvastatin, metformin, glipizide glimepiride cocktail using high throughput CYP450 enzyme inhibition assay; against CYP1A2, 2C19, 2D6 and 3A4 isozymes. Results Contents of arjunolic acid, arjunetin, berberine, piperine, resveratrol and withaferin-A in Ridayarishta formulation were found to be 1.76±0.12, 1.51±0.09, 1.85±0.05, 3.2±0.12, 1.21±0.08, and 2.16±0.09 ppm, respectively. Quantity of ethanol in Ridayarishta was found to be 7.95±0.023% (V/V). Ridayarishta showed significantly higher (P<0.001) IC 50 value against CYP1A2 (IC 50 –13.80±1.96 µg/mL), 2C19 (IC 50 –14.343±2.28 µg/mL), 2D6 (IC 50 –0.897±0.28 µg/mL) and 3A4 (IC 50 –32.057±2.51 µg/mL) compared to positive controls such as furafylline, tranylcypromine, quinidine and ketoconazole respectively. Cocktail of herbal formulation and cardio protective, antihypertensive, anti-diabetic drugs showed significantly (P<0.001and P<0.01) less or negligible HDI. Conclusion Ridayarishta formulation alone and cocktail with amlodipine besilate, atenolol, atorvastatin, metformin, glipizide, glimepiride had negligible or insignificant effect on CYP450 inhibition. It may be concluded that consumption of Ridayarishta along with selective cardio protective, antihypertensive and anti-diabetic conventional medicine is safe with negligible or without any significant CYP450 (CYP1A2, 2C19, 2D6 and 3A4) inhibition mediated HDI. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Heinsia crinita is used in traditional medicine for the treatment of febrile illness and erectile dysfunction. Its stem bark powder is found in some peripheral markets in the Democratic Republic of the Congo (DRC) as a remedy against malaria. Investigations were conducted on crude extracts of leaves, fruits and stem barks in view to validate their use and to determine which plant part possesses the best antiplasmodial properties. Materials and methods Different plant parts were extracted with methanol, ethanol and dichloromethane. Based on the preliminary assays, the dichloromethane extract of the stem bark was subjected to fractionation using preparative HPLC system and column chromatography. This step led to the isolation of two new iridoids which had their structures elucidated by NMR, UV, MS and FT-IR spectroscopic techniques. Extracts and pure compounds were tested in vitro against the 3D7 strain of Plasmodium falciparum. The inhibition of the parasite growth was evaluated in vitro by colorimetric method ( p -LDH assay) and their cytotoxicity evaluated in vitro against the human non-cancer fibroblast cell line (WI38) through WST1 assay. The in vivo antiplasmodial activity was assessed by the inhibition of Plasmodium berghei growth in infected mice treated with the ethanol extract of H. crinita stem bark at the concentrations of 200 and 300 mg/Kg/day per os , using a protocol based on the 4-d suppressive test of Peters and compared to a non-treated negative control group of mice (growth =100%). Finally the antioxidant activity of the same extract was evaluated using ABTS, DPPH and cell-based assays. Results A moderate in vitro antiplasmodial activity was observed for the dichloromethane extract of the stem bark of H. crinita (IC 50 =29.2±1.39 µg/mL) and for the two new iridoids, lamalbide 6, 7, 8- triacetate (IC 50 =16.39±0.43 µg/mL) as well as for its aglycone lamiridosin 6, 7, 8-triacetate (IC 50 =0.44.56±1.12 µg/mL). The ethanolic stem bark extract (200 and 300 mg/kg/day, oral route) showed a moderate in vivo antimalarial activity in Plasmodium berghei -infected mice with 27.84±2.75% and 48.54±3.76% of inhibition of the parasite growth, respectively (p<0.01).). This extract displayed high cellular antioxidant activity using dichlorofluorescein-diacetate (DCFDA) on HL-60 monocytes. These crude extracts and pure compounds tested at the higher concentration of 100 µg/mL did not show any cytotoxicity against WI38 cells. Conclusions The results showed that H. crinita extracts possess antimalarial activity and contain some unusual iridoids with moderate antiplasmodial activity, therefore justifying to some extent its traditional use by the local population in DRC for this purpose. This is the first report of the isolation and antiplasmodial activity of these two new iridoids. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Hedyotis diffusa is an ethno-medicine used for anti-cancer treatment in the clinic of traditional Chinese medicine (TCM). The total coumarins of Hedyotis diffusa (TCHD) was a selected extract with observed antiproliferative activity, which has not been tested in treatment of myelodysplastic syndromes (MDS) or acute myeloid leukemia (AML). Aim of the study This study aimed to evaluate the apoptosis-inducing effect of TCHD on human MDS cell line (SKM-1) and explore its action mechanism in association with caspase family and PI3K/Akt signaling pathway. Materials and methods The chemical constituents and total coumarins content of TCHD were determined by High Performance Liquid Chromatography-tandem mass spectrometry (HPLC-MS/MS) and UV–vis spectrophotometry, respectively. MTT assay, Hoechst 33258 staining, and Annexin V-FITC/PI double labeling were applied to evaluate TCHD's efficacy on SKM-1 cells. Western blot analysis was also used to clarify the action mechanism of TCHD on protein expression level. Results Two compounds, p -coumaric acid and E -6-O- p -coumaroyl scandoside methyl ester, were identified in TCHD, and its total coumarins content reached 87.4%. By MTT assay, apoptosis-inducing effect of TCHD on SKM-1 cells was found in a dose-dependent manner after 24–48 h treatment, with IC 50 values of 104.48 μg/ml and 100.66 μg/ml, respectively. Morphological and flow cytometry observation also confirmed such effect of TCHD. Western blot analysis clarified its action mechanism associating with the activation of caspases and inhibition of PI3K/Akt pathway proteins. Conclusions This is the first report regarding the apoptosis-inducing efficacy and mechanism of TCHD on SKM-1 cells, providing a promising candidate of TCM for MDS and AML therapy with fewer side effects. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Bullatine A, a C 20 -diterpenoid alkaloid and one of the major effective ingredients in Aconiti brachypodi Radix (Xue-shang-yi-zhi-hao), can block pain hypersensitivity in a variety of rodent models through expression of spinal microglial dynorphin A. Aim of the study To assess the interaction between bullatine A and morphine on antinociception in acute nociception and pain hypersensitivity states, with the exogenous synthetic dynorphin A as a comparison Materials and methods Spinal nerve ligation-induced neuropathic rats and naïve mice were used for assessing the acute and chronic interactions of bullatine A/dynorphin A with morphine. Results Single subcutaneous injection of bullatine A or dynorphin A(1−17) did not either alter formalin- and thermally (hot-plate and water immersion tests)-induced acute nociception or potentiate morphine antinociception in naïve mice. In contrast, bullatine A dose-dependently inhibited formalin-induced tonic pain with the efficacy of 54% inhibition and the half-effective dose of 0.9 mg/kg. Concurrent bullatine A additively enhanced morphine antinociception. In neuropathic rats, the antinociceptive effects of multiple bidaily intrathecal injections of bullatine A and dynorphin A remained consistent over 13 days, whereas morphine produced progressive and complete tolerance to antinociception, which was completely inhibited by concurrent bullatine A and dynorphin A. A single intrathecal injection of bullatine A and dynorphin A immediately reversed established morphine tolerance in neuropathic rats, although the blockade was a less degree in the thermally induced mouse acute nociceptive tests. The inhibitory effects of bullatine A and dynorphin A on morphine tolerance were immediately and completely attenuated by intrathecal dynorphin A antibody and/or selective κ-opioid receptor antagonist GNTI. Conclusion These results suggest that bullatine A produces antinociception without induction of tolerance and inhibits morphine antinociceptive tolerance, and provide pharmacological basis for concurrent bullatine A and morphine treatment for chronic pain and morphine analgesic tolerance. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Houttuynia cordata is an important traditional Chinese medicine used in heat-clearing and detoxifying, swelling and discharging pus, promoting diuresis and relieving stranguria which recorded in Pharmacopoeia of the people's Republic of China (2015 Edition). H. cordata has been recorded in the book Bencaogangmu which was written by Shizhen Li for the treatment of pyretic toxicity, carbuncle swelling, haemorrhoids, and rectocele diseases. Aim of the study Phytochemical investigation of the aerial parts of H. cordata and evaluation of their PTP1B inhibitory activities and hepatoprotective activities. Materials and methods The dried aerial parts of H. cordata were fractionated by liquid-liquid extraction to obtain CHCl 3 , ethyl acetate, and n-butanolic fractions. The CHCl 3 fraction was confirmed active fraction by the bioactivity-guided investigation, which was isolated and purified by chromatographing over silica gel, Sephadex LH-20, MPLC, and preparative HPLC. The chemical structures of the purified compounds were identified by their spectroscopic data and references. Results Eight new compounds ( 1 – 8 ), together with fourteen known compounds ( 9–22 ) were isolated from the aerial parts of H. cordata. The known compounds ( 9–22 ) were obtained from this plant for the first time. Among them, some compounds exhibited moderate bioactivities. Conclusion Compounds ( 1–8 ) were identified as new alkaloids, and the known alkaloids ( 9–22 ) were isolated from this plant for the first time. Compounds 1 , 4 , 14 , and 19 showed significant PTP1B inhibitory activities with IC 50 values of 1.254, 2.016, 2.672, and 1.862 µm, respectively. Compounds 1 , 3 , 6 , 11 , 17 , and 20 (10 µm) exhibited moderate hepatoprotective activities against D-galactosamine-induced WB-F344 cells damage. [ABSTRACT FROM AUTHOR]

Objective To assess the cumulative evidence on the efficacy and effectiveness of Ginkgo biloba extract (GbE) in the treatment of dementia. Design Overview of systematic reviews. Methods PubMed/MEDLINE, EMBASE, Cochrane, and Google Scholar were searched in June 2016. Systematic reviews (SRs) of randomized controlled trials (RCTs) evaluating the effects of GbE on different outcomes in people with dementia or cognitive impairment were included. Methodological quality of the included SRs was assessed using the AMSTAR tool. The quality of evidence of the primary studies was assessed using GRADE. Results Twelve SRs with meta-analyses met the eligibility criteria. The quality of the evidence reported in these SRs varies ranging from low to moderate level. Overall, the available evidence suggests that GbE has potentially beneficial effects over placebo on cognitive performance, activities of daily living, and clinical global impression in the treatment of dementia at doses greater than 200 mg/day (usually 240 mg/day) administrated for 22 weeks or longer, and that GbE appears to be safe for human consumption. No sufficient evidence supports the favorable effects of GbE administrated for less than 22 weeks. The available evidence consistently indicates that a dose less than 200 mg/day of GbE may not be adequate to yield clinical relevant effects in the treatment of dementia. Conclusions GbE has potentially beneficial effects for people with dementia when it is administered at doses greater than 200 mg/day for at least 5 months. Given the lower quality of the evidence, further rigorously-designed, multicenter-based, large-scale RCTs are warranted. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Pueraria lobata root was used to treat wasting-thirst regarded as diabetes mellitus and was included in the composition of Okcheonsan, which is prescribed for thirst-waste in traditional Chinese medicine. Aim of the study The objective of this study was to evaluate the anti-diabetic potential of the root of Pueraria lobata and its constituents via protein tyrosine phosphatase 1B (PTP1B) and α-glucosidase inhibitory activities. Materials and methods In this study, anti-diabetic activities of the 70% ethanolic (EtOH) extract from P. lobata roots and its solvent soluble fractions with the isolated compounds were investigated by evaluating in vitro PTP1B and α-glucosidase inhibitory activities. We also examined the potentials of active compounds as PTP1B and α-glucosidase inhibitors via enzyme kinetics and in silico molecular docking simulation between the enzymes and active compounds. Results Triterpenoids lupeol and lupenone were potent PTP1B inhibitors with IC 50 values of 38.89±0.17 and 15.11±1.23 μM. Kinetic study using the Lineweaver-Burk and Dixon plots demonstrated that these compounds showed a noncompetitive-type inhibition against PTP1B with respective K i values of 13.88 μM and 21.24 μM. In addition, molecular docking simulation showed lupeol and lupenone has negative binding energy values of −8.03 and −8.56 kcal/mol. Considering the α-glucosidase inhibitory potential, daidzein, genistein, and calycosin exhibited the most potent α-glucosidase inhibition with IC 50 values of 8.58±0.94, 2.37±0.52 and 6.84±1.58 μM, respectively. Kinetic study demonstrated that these 3 compounds showed a noncompetitive-type inhibition against α-glucosidase with respective K i values of 17.64 μM, 5.03 μM and 13.83 μM. Moreover, molecular docking simulation showed daidzein, genistein and calycosin has more lower binding energy (−7.16 kcal/mol, −7.42 kcal/mol and −7.31 kcal/mol) with higher binding affinity and tight binding capacity in the molecular docking studies than standard ligand α-D-glucose (−6.74 kcal/mol). Conclusion Our results of the present study clearly demonstrate the potential of P. lobata extract and its constituents to inhibit PTP1B and α-glucosidase, contributing to the development of therapeutic or preventive agents that can be used in the treatment of diabetes. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance The Natal gwarri or Natal ebony ( Euclea natalensis A.DC.) is a deciduous tree found widespread throughout southern Africa, especially in Kwazulu-Natal and the southern cost. It has been widely used by indigenous communities such as the Zulus, Tsongas and Vendas for symptoms related to tuberculosis (TB). The decoctions made from the plant parts are administered for chest diseases to treat complications such as chest pains, bronchitis, pleurisy and asthma. TB is prevalent in immune-compromised patients and it is evident that TB-drugs cause hepatotoxicity. The objective of the present study was therefore to evaluate the antimycobacterial activity of the ethanolic extract of E. natalensis against TB and its hepatoprotective and immunomodulatory activities. Materials and methods The antimycobacterial, antioxidant, hepatoprotective, immunomodulatory activity and cytotoxicity of the ethanolic extract of the shoots of E. natalensis were determined in vitro . The mechanism of action of the antituberculosis activity was determined by investigating the inhibitory effect on mycothiol disulfide reductase enzyme. Furthermore, the acute, sub-acute toxicity (50–2000 mg/kg) and antimycobacterial effect (300 mg/kg) of E. natalensis shoot extract were investigated in Balb/c mice. Hepatoprotective activity of the extract (50–150 mg/kg) was evaluated on isoniazid and rifampicin (50 mg/kg; i.p.) induced hepatic damage in a rat model. Results The minimum inhibitory concentration of the extract was found to be 125 µg/ml against Mycobacterium tuberculosis . The extracts 50% inhibitory concentration (IC 50 ) against 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical was found to be 22.55 µg/ml. The plant showed a hepatoprotective effect (50% at 12.5 µg/ml) and the ability to increase T-helper 1 cell cytokines; Interleukin 12, Interleukin 2 and Interferon α by up to 12 fold and the ability to decrease the T-helper 2 cell cytokine Interleukin 10 4 fold when compared to baseline cytokine production. No cellular toxicity was observed in primary peripheral blood mononuclear cells (PBMC's) and two secondary cell lines; U937 monocytes and Chang liver cells (a derivative of the HepG2 cell line). During mechanistic studies, the extract showed a 50% inhibition of mycothiol reductase activity at 38.62 µg/ml. During the acute and sub-acute studies, E. natalensis exhibited no toxic effect and the 50% lethal dose (LD 50 ) was established to be above 2000 mg/kg. The extract was able to reduce the mycobacterial load (1.5-fold reduction) in infected mice. Isoniazid and rifampicin caused significant hepatic damage in rats, and the extract was able to reduce the toxicity by 15% and 40% at 50 and 150 mg/kg respectively. Conclusion The present study supports the traditional usage of the plant against tuberculosis symptoms. The study showed the ability of E. natalensis shoot extract to inhibit mycobacterial growth, stimulate an appropriate immune response and have a hepatic protective effect. Due to the extract's significant results for hepatoprotective, immunomodulatory effects and antimycobacterial activity, it may prove to be effective to serve as an adjuvant for TB-patients. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Calotropis gigantea (L.) Dryand (Apocynaceae) is a medicinal plant native to southern China, India and Southeast Asia. It has been traditionally used for the treatment of several diseases including cancers in these countries. Aim of the study This study aimed to isolate bioactive cardenolides from C. gigantea , to screen their hypoxia-inducible factor (HIF-) 1 inhibitory activity, and to analyze the structure-activity relationship (SAR). Materials and methods Isolation and purification of cardenolides from the latex and the fruits of C. gigantea were performed by using a series of separation techniques. Their structures were fully characterized by elucidating their NMR and HRMS data. The HIF-1 inhibitory activities of cardenolides were evaluated by using a T47D cell-based dual-luciferase reporter assay. The potent cardenolides were selected to further evaluate their dose-response manner. Cytotoxic effects of selected cardenolides were also examined against breast cancer cell line (MCF-7) and normal mammary epithelial cell line (MCF-10A) by MTT assay. Results Among twenty isolated cardenolides, compounds 1 , 3 , 4 , 6–8 , 14 and 17 exhibited stronger HIF-1 inhibitory activities than that of digoxin, a well-known HIF-1 inhibitor ( P <0.001). These eight cardenolides inhibited HIF-1 transcriptional activity in a dose-dependent manner with IC 50 values in nanomolar potency (21.8–64.9 nM). An analysis of SAR revealed the great contributions of a β -configuration of the substituents at positions of C-2′ and C-3′, an aldehydic moiety on C-19, and the dioxane moiety between the aglycone and sugar parts of cardenolides to the HIF-1 inhibitory activity. In contrast, a hydroxyl group at any positions of C-15, C-16 and C-4′ of cardenolides showed negative effects on suppressing HIF-1 transcriptional activity. In addition, these eight cardenolides also exhibited potent cytotoxic effects against human breast cancer cell MCF-7 (IC 50 values ranged from 30.5 to 68.8 nM), but less toxic effects to human normal mammary epithelial cell MCF-10A (IC 50 values >20 µM). Conclusions This is the first report of a comprehensive study of SAR on cardenolides from C. gigantea as HIF-1 inhibitors. Eight cardenolides ( 1 , 3 , 4 , 6–8 , 14 and 17 ) showed both potent HIF-1 inhibitory activity and strong cytotoxic effect against MCF-7 cancer cells in nanomolar level. The findings of these cardenolides provided important insights into the development of these potent HIF-1 inhibitors as anticancer drug. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Styrax liquidus is a resinous exudate (balsam) obtained from the wounded trunk of the Liquidambar orientalis Mill. (Hamamelidaceae). Styrax has been used for treatment of various ailments in Turkish folk medicine such as skin problems, peptic ulcers, nocturnal enuresis, parasitic infections, antiseptic or as expectorant. Aim of study In spite of frequent use of styrax in Turkish folk medicine as well as once as a stabilizer in perfumery industry, negative reports have been noticed by the international authority for restriction its use based on some limited evidences from an in vitro study. The aim of the present study was to evaluate the genotoxic and cytotoxic potential of styrax and its ethanolic extract using in vivo and in vitro assays, as well as an antimutagenic assay and also to determine its phenolic constituents with chromatographic analysis. Materials and methods In vitro mutagenicity and antimutagenicity of styrax and its ethanolic extract were evaluated by Ames test performed on Salmonella TA98 and TA100 strains with and without metabolic activation (10– 30,000 µg/plate). The genotoxicity was also studied in vivo by chromosomal aberrations assay on bone marrow of Balb C mice with different its concentrations (500–2000 mg/kg body weight). Cytotoxicity has been evaluated by the MTT assay using L929 cell line. Its phenolic constituents were determined by HPLC analysis. Results Genotoxicological investigations of styrax or its ethanolic extract showed that none of the tested concentrations induced a significant increase in the revertant number of TA98 and TA100 strains with or without metabolic activation, indicating no mutagenicity to the tested strains. Also results indicated that up to 2000 mg/kg body weight, styrax is not genotoxic in mammalian bone marrow chromosome aberration test in vivo . In cytotoxicity study, the IC 50 values of styrax and its ethanolic extract were found to be 50.22±1.80 and 59.69±11.77 µg/mL, respectively. Among the studied reference standards the major phenolic acids in styrax balsam was found to be p-coumaric acid (2.95 mg/g), while in its ethanolic extract not only p-coumaric acid (11.46 mg/g), but also gallic acid (1.60 mg/g) were found to the main components. Conclusion The findings of the present study provide scientific basis to the safety of styrax from the viewpoint of genotoxicity risk, and in fact, it was found to be beneficial against genotoxicity. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance The rhizomes of Dioscorea membrancea Pierre have been used in Thai traditional medicine as an ingredient formula for liver cancer and cholangiocarcinoma treatment. Aim of the study To investigate the cytotoxic activity of ethanolic extract and constituents of D. membrancea to support its traditional use. Materials and methods The SRB assay was used to determine the cytotoxic activity against hepatocellular carcinoma (HepG2), cholangiocarcinoma (KKU-M156) cells and one normal human keratinocyte immortal cells (HaCaT) with its ethanolic extract and isolated compounds. Bioassay guided isolation was used for isolating cytotoxic compounds. Results The ethanolic extract of D . membranacea rhizome showed weak cytotoxic against KKU-M156 and HepG2 (IC 50 at 72 h exposure=30.49±0.82 and 38.97±2.04 µg/mL respectively). A new steroid [epipanthogenin B ( 1 )], a known steroid [panthogenin B ( 2 )], two napthofuranoxepins [dioscorealide A ( 3 ) and dioscorealide B ( 4 )], phenanthraquinone [dioscoreanone ( 5 )] and two phenanthrene [5,6-dihydroxy-2,4-dimethoxy-9,10-dihydrophenanthrene ( 6 ) and 2,5,6-trihydroxy-3,4-dimethoxy, 9, 10-dihydrophenanthrene ( 7 )] were isolated from active chloroform fraction. Compound 4 showed the highest cytotoxicity against HepG2 (IC 50 at 72 h exposure=2.87±0.21 µM) and KKU-M156 (IC 50 at 72 h exposure=1.67±0.10 µM) and less toxicity against normal cell line (HaCaT) (IC 50 at 72 h exposure>100 µM). Compound 5 showed selective cytotoxic activity against KKU-M156 (IC 50 at 72 h exposure=3.46±0.11 µM). Compounds 6 and 7 showed weak cytotoxic activity against HepG2 (IC 50 at 72 h exposure=24.96±2.32 and 51.31±3.52 µM). Compounds ( 1 – 3 ) showed no cytotoxic activity against HepG2 and KKU-M156 cell lines (IC 50 at 72 h exposure>100 µM). Conclusion Seven compounds were isolated from active chloroform fraction of the ethanolic extract of D . membranacea rhizomes. Only dioscorealide B ( 4 ) might be served as a good anticancer agent for liver cancer and cholangiocarcinoma cancer because it can kill cancer cell but not toxic on normal cell. This research support Thai traditional medicine use of D. membranacea for liver cancer and cholangiocarcinoma cancer treatment. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance: Coronopus didymus Linn. (Brassicaceae) is a medicinal plant used traditionally as antipyretic, expectorant, to purify blood and for alleviating symptoms of pain, inflammations, malaria, wounds and cancer. Aim of the study: The present study was designed to isolate and identify the cytotoxic compounds responsible for anticancer activity from this traditionally useful medicinal plant. Materials and methods: Bioassay-guided fractionation of the ethanolic extract of aerial parts of C. didymus allowed the isolation of compounds responsible for anticancer activity. Their structures were elucidated by UV Spectroscopy (with shift reagents), ESI-MS and NMR spectral data. Preliminary anticancer activity of ethanolic extract, different fractions and isolated compounds was assessed through MTT in vitro cytotoxicity assay in a dose dependent manner against human cancer cell lines (HeLa and LN18) and normal 293T cells. Results: Three flavonoids namely 5,7,4′-trihydroxy-3′-methoxyflavone-4′- O - β - D -glucoside ( 1 ), 5,7,4′-trihydroxy-3′-methoxyflavone-4′- O -(6''-acetyl)- β - D -glucoside ( 2 ) and 5,7,4′-trihydroxy-3′-methoxy flavone ( 3 ), were isolated from aerial parts. Compound 1 was identified for the first time from the genus Coronopus . All the compounds 1 – 3 showed promising activity against HeLa cells with IC 50 values of 43.50, 0.63 and 3.67 µM , respectively. Significant result was also obtained with compound 3 against LN18 cells with IC 50 value of 46.63 µM . Conclusion: The cytotoxic activity of the crude extract and fractions which may largely be due to its major isolated constituents, flavonoids 1 – 3 , against both HeLa and LN18 cells provides a scientific basis for the ethnopharmacological use of C. didymus as anticancer agent. [ABSTRACT FROM AUTHOR]