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Summary Introduction Carica papaya L. belongs to Caricaceae family. It is an evergreen tree or shrub with usually unbranched trunk. Previous studies on the methanolic leaf extract of this plant proved the antibacterial, cytotoxic, anticancer and antioxidant effects indicative of promising therapeutic potentials. Methods Chromatographic and spectroscopic techniques along with high performance liquid chromatography quantitative analysis were performed to isolate, tentatively identify and quantify polyphenolic compounds in MeOH extract of C. papaya leaves. UHPLC-DAD-ESI-MS/MS and UHPLC-DAD methods were adopted for qualitative and quantitative analysis respectively. In the case of polyphenol separation, some reference substances were isolated by preparative HPLC. Results Six polyphenols were isolated and quantified from C. papaya leaf extract, including three phenolic acids: derivatives of caffeic acid, p-coumaric acid, ferulic acid and three flavonoids: quercetin dirhamnosylhexoside, kaempferol dirhamnosyl-hexoside, quercetin 3-O-rutinoside. Among them quercetin dirhamnosyl-hexoside was significantly higher (1.9 mg/g extract) than other identified compounds. Conclusion The results of present study provide a new data on polyphenols composition of C. papaya plant derived from Uganda and promote research on its possible standardization and use as a dietary supplement and/or herbal medicine.

Given the strong associations between diet and cancer risk, there is considerable scientific interest in determining whether dietary factors associated with prostate cancer cell implantation may influence epigenetic alternations. The aim of the research was to assess impact of selected trace elements (selenium, zinc and copper) on the kinetics of changes (10-13-14-21 week of life cycle of rats) in the level of 7-methylguanine, 3-methyladenine, 1-methylguanine and 8-oxo-guanine in the urine of rats with implanted prostate cancer cells (LNCaP). Modified nucleobases were determined by validated high performance liquid chromatography coupled to mass spectrometry (LC-MS/MS) method using multiple reaction monitoring (MRM) mode. In the presented model the implantation of rats with cancer cells did not affect the level of the examined biomarkers in the rats’ urine. The level of methyl derivatives was statistically significantly reduced with the age of the examined rats. The implantation of rats with cancer cells results in the appearance of tumors in 71% of the rats obtaining the standard diet and respectively in 25% of those supplemented with selenium. Supplementation with selenium affects both the effectiveness of tumor induction and the concentration of 7-MeG, 3-MeA, 1-MeG and 8-oxoG in urine of the examined rats. These findings show that modified nucleosides can play an important role in cancer prevention.

Sigesbeckia orientalis, more commonly referred to as Herba Sigesbeckiae or Xi Xian Cao in traditional Chinese medicine and hy thiêm in traditional Vietnamese medicine, is used in China and Vietnam to treat inflammatory diseases such as arthritis, rheumatism, and joint pain. In initial investigations, the dichloromethane extract from the aerial parts of S. orientalis showed distinct inhibitory effects on the release of interleukin-8 in human neutrophils. Therefore, the purpose of the present study was the phytochemical investigation of the bioactive dichloromethane extract and the in vitro analysis of the effects of the isolated compounds on interleukin-8, interleukin-1β, tumor necrosis factor-α, and monocyte chemoattractant protein 1 release, and surface expression of adhesion molecules (CD11a, CD11b, and CD62L) in lipopolysaccharide-stimulated human neutrophils to identify the active principle(s). The separation of the bioactive dichloromethane extract using various chromatographic techniques led to the isolation of nine compounds. Their chemical structures were elucidated from nuclear magnetic resonance and mass spectrometry data. One diterpene, 17(13 → 14)-abeo-ent-3S*,13S*,16-trihydroxystrob-8(15)-ene, was identified as a new natural product. Three germacranolide sesquiterpene lactones inhibited interleukin-8 production with IC50 values between 1.6 and 6.3 µM, respectively, and tumor necrosis factor-α production with IC50 values between 0.9 and 3.3 µM, respectively. Furthermore, they significantly inhibited interleukin-1β and monocyte chemoattractant protein 1 production and diminished the effects of lipopolysaccharide on the surface expression of the adhesion molecules CD11a, CD11b, and CD62L. These findings support the traditional use of S. orientalis in the treatment of inflammatory diseases.

Carpesium divaricatum Sieb. &amp, Zucc., a traditional medicinal plant used as an inflammation-relieving remedy, is a rich source of terpenoids. At least 40 germacrane-type sesquiterpene lactones, representatives of four different structural groups, were isolated from the plant. Cytotoxicity against cancer cells in vitro is the most frequently described biological activity of the compounds. However, little is known about the selectivity of the cytotoxic effect. The anti-inflammatory activity of the germacranolides is also poorly documented. The objective of the present study was to assess the cytotoxic activity of selected C. divaricatum germacranolides-derivatives of 4,5,8,9-tetrahydroxy-3-oxo-germacran-6,12-olide towards cancer and normal cell lines (including cells of different p53 status). Moreover, to assess the anti-inflammatory effect of the compounds, the release of four proinflammatory cytokines/chemokines (IL-1β, IL-8, TNF-α and CCL2) by lipopolysaccharide-stimulated human neutrophils was measured by ELISA. The investigated sesquiterpene lactones demonstrated nonselective activity towards prostate cancer (Du145 and PC3) and normal prostate epithelial cells (PNT2) as well as against melanoma cells (A375 and HTB140) and keratinocytes (HaCaT). Cytotoxic activity against osteosarcoma cells was independent of their p53 status. In sub-cytotoxic concentrations (0.5–2.5 µM) the studied compounds significantly decreased cytokine/chemokine release by lipopolysaccharide-stimulated human leukocytes.

Carpesium divaricatum Sieb. & Zucc., a traditional medicinal plant used as an inflammation-relieving remedy, is a rich source of terpenoids. At least 40 germacrane-type sesquiterpene lactones, representatives of four different structural groups, were isolated from the plant. Cytotoxicity against cancer cells in vitro is the most frequently described biological activity of the compounds. However, little is known about the selectivity of the cytotoxic effect. The anti-inflammatory activity of the germacranolides is also poorly documented. The objective of the present study was to assess the cytotoxic activity of selected C. divaricatum germacranolides-derivatives of 4,5,8,9-tetrahydroxy-3-oxo-germacran-6,12-olide towards cancer and normal cell lines (including cells of different p53 status). Moreover, to assess the anti-inflammatory effect of the compounds, the release of four proinflammatory cytokines/chemokines (IL-1β, IL-8, TNF-α and CCL2) by lipopolysaccharide-stimulated human neutrophils was measured by ELISA. The investigated sesquiterpene lactones demonstrated nonselective activity towards prostate cancer (Du145 and PC3) and normal prostate epithelial cells (PNT2) as well as against melanoma cells (A375 and HTB140) and keratinocytes (HaCaT). Cytotoxic activity against osteosarcoma cells was independent of their p53 status. In sub-cytotoxic concentrations (0.5–2.5 µM) the studied compounds significantly decreased cytokine/chemokine release by lipopolysaccharide-stimulated human leukocytes.

Introduction In European traditional medicine, common ash leaf infusion is recommended by European Medicines Agency to treat minor articular pain and to increase the amount of urine for flushing minor urinary complaints. However, a comprehensive ultra-high-performance liquid chromatography diode array detector electrospray ionisation tandem mass spectrometry (UHPLC-DAD-ESI-MS/MS) analysis of this pharmacopeial plant material has never been performed. Moreover, the number of biological and pharmacological investigations proving the usefulness of this plant material in recommended traditional uses is surprisingly small. Objective Phytochemical profiling of ash leaf samples from different commercial and natural sources and the determination of the in vitro effects on inflammatory mediators in a model of human neutrophils. Methods Ash leaf samples were characterised by total hydroxycinnamic acid content and by high-performance thin layer chromatography (HPTLC), UHPLC-DAD-ESI-MS/MS methods. The effects of leaf infusions on reactive oxygen species (ROS), tumor necrosis factor (TNF-α), interleukin 8 (IL-8), interleukin 1β (IL-1β), and monocyte chemoattractant protein 1 (MCP-1) production by neutrophils were measured using luminol-dependent chemiluminescence and enzyme-linked immunosorbent assay (ELISA). Results In ash leaf samples 64 compounds were identified or partly identified together with four unknown compounds. The major compounds detected belong to different structural groups, including phenolic acid derivatives, phenylethanoids, flavonoids, iridoids, secoiridoids and lignans. The major compounds detected in ash samples were chlorogenic acid, quercetin-3-O-rutinoside, verbascoside, oleuropein and ligstroside. However, one sample contained coumarin derivatives. This finding suggested adulteration with other Fraxinus species and/or plant parts. All infusions were able to inhibit ROS, cytokine and chemokine production. Conclusions The performed phytochemical and biological analyses contribute to the knowledge about this pharmacopeial plant material and supports its traditional use to treat minor inflammatory complaints.

In traditional European medicine, common lilac (Syringa vulgaris L., Oleaceae) bark has been used in the form of an infusion, decoction or alcoholic extract as an antipyretic and a cold and cough treatment. Our primary study showed that alcoholic extract of the bark was able to decrease the release of interleukin- 8 in an in vitro model The aim of the study was to investigate the phytochemical composition of common lilac bark extract by the HPLC-DAD-MS/MS method, isolate the dominant compounds and investigate their effects on the proinflammatory functions of human neutrophils and monocytes/macrophages. Syringin (sinapyl alcohol-O-glucoside), olivil-4′-O-glucoside, sinapyl aldehyde-O-glucoside and forsythoside B were isolated using RP-18, Sephadex LH-20 column chromatography and preparative chromatography. The structures were confirmed by the UV, MS and 1H and 13C NMR spectra. The effects on cytokine (TNF-α, IL-1β, IL-6) and chemokine (IL-8 and MCP-1) production by neutrophils and monocytes/macrophages were measured using an enzyme-linked immunosorbent assay (ELISA). Moderate anti-inflammatory activity was observed in the neutrophil model for all compounds compared with oleuropein, a common compound in the Oleaceae family with well-documented anti-inflammatory effects. However, syringin at concentration of 50 μM significantly inhibited TNF-α production and stimulated TGF-β release in LPS-treated monocytes/macrophages. Our results demonstrated that common lilac bark is a valuable source of active compounds, especially syringin.

Different in vitro cultures of the Turkish origin medicinal plant species, Salvia viridis, were estimated in order to investigate their potential for the accumulation of phenolic compounds. Among the seventeen detected compounds, thirteen phenolic acid derivatives and four phenylethanoids were identified in extracts from shoot and/or callus cultures using the UPLC-DAD/ESI-MS method. The accumulation of the compound depended on the type of culture (callus, shoot), the medium composition and age of culture (6, 12, 18 and 24 months in in vitro conditions). Six of the seventeen detected compounds were estimated in all the analyzed samples, however, sometimes only in trace amounts. The predominant metabolite was rosmarinic acid (RA). Its content ranged between 2.4 and 11.5 mg/g dry weight (DW). The highest levels of rosmarinic acid and total phenolic compounds were found in a half-year-old shoot culture. The content of the RA was nine times higher than that in the shoots of plant grown in field conditions. The antioxidant activity of S. viridis cultures was examined by the O2− and 1-1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging test and FRAP (ferric reducing antioxidant power) assay. In addition, the antifungal and the antibacterial activities were examined against a wide spectrum of micro-organisms. All the samples showed promising antioxidant potential; however, the shoot extract had the highest antioxidant activity in all the assays. Preliminary results demonstrated that all the analyzed extracts inhibited the growth of Gram-positive and Gram-negative bacteria. The extracts also showed strong activity against Candida and Aspergillus species.

Background Among solar radiation, ultraviolet light is the most harmful for the skin, because of intracellular reactive oxygen species formation, leading to oxidative stress, cell damage and apoptosis. Crucial role in skin protection against oxidative stress play antioxidant enzymes regulated by Nrf2 transcription factor. Some plant-derived polyphenols are known to protect skin fibroblasts against UV through induction of Nrf2-dependent antioxidant genes expression. Purpose We previously found out that water extracts from Galinsoga sp. herb protected human dermal fibroblasts against UVA-induced oxidative stress and apoptosis. However, which compounds were responsible for such protective action remained unclear. Here, we investigated photoprotective potential and mechanism of action of two main isolated compounds, 2,3,5(2,4,5)-tricaffeoylaltraric acid and 2,4(3,5)-dicaffeoylglucaric acid, on human dermal fibroblasts (NHDF). Study design/Methods NHDF cells were pretreated with tested compounds (6.25–50 µM) and irradiated with UVA (25 J/cm2). Intracellular ROS and GSH level, cell viability, cell membrane integrity and apoptosis were measured. HO-1 protein expression and Nrf2 transcription factor activation were also assessed. Results Cells pretreated with tested compounds prior to UVA showed inhibition of intracellular ROS formation and increase of GSH level. Significant increase of cell viability was also observed, as well as decrease of LDH release and a the rate of apoptotic cells in comparison to untreated cells. Furthermore, tested compounds increased HO-1 expression and activated the Nrf2 transcription factor in NHDF cells. Conclusion Present study demonstrated that caffeic acid derivatives present in Galinsoga parviflora herb, in particular tricaffeoylaltraric acid may protect dermal fibroblasts against UVA-induced oxidative stress through activation of intracellular antioxidative system. Such caffeic acid derivatives are bioactive compounds which might prevent UV-induced photoageing and photocarcinogenesis.

Plants have been used for medical purposes since ancient times. However, a detailed analysis of their biological properties and their associated active compounds is needed to justify their therapeutic use in modern medicine. The aim of the study was to identify and quantify the phenolics present in hydromethanolic extracts of the roots and shoots of the Chinese Salvia species, Salvia bulleyana. The qualitative and quantitative analyses were carried out by ultrahigh-performance liquid chromatography with electrospray ionization mass spectrometry detection (UHPLC-PDA-ESI-MS), and high-performance liquid chromatography with photodiode array (HPLC-PDA) detection. The extracts of S. bulleyana were also screened for their antioxidant activity using ferric ion (Fe3+) reducing antioxidant power (FRAP), 1,1-diphenyl-2-picrylhydrazyl (DPPH), diammonium 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) cation (ABTS), superoxide radical anion (O2•–), and inhibition of lipid peroxidation assays. The S. bulleyana extracts were found to contain 38 substances, of which 36 were phenols, with a total level of 14.4 mg/g DW (dry weight) in shoots, and 23.1 mg/g DW in roots. Twenty-eight phenols were polyphenolic acids or their derivatives, the most abundant in shoots being rosmarinic acid, and in roots, salvianolic acid K followed by rosmarinic acid. The other major phenolic acids were caffeic acid, caffeoyl-threonic acids, isomers of lithospermic acid, salvianolic acid F, salvianolic acid B, and yunnaneic acid E. In addition to polyphenolic acids, nine flavonoids were detected in the shoot extract. While both extracts showed significant antioxidant activity, the shoot extract, containing both polyphenolic acids and flavonoids, demonstrated a slightly greater antioxidant potential in some of the anti-radical tests than the roots. However, the root extract proved to be slightly more effective in the lipid peroxidation inhibition test. Thus, S. bulleyana was demonstrated as a promising source of antioxidants, and worthy of further more detailed studies.

Extracts from the roots of Ononis spinosa L. (restharrow roots) are traditionally used for the treatment of patients with urinary tract infections due to its mild diuretic activity, caused by the inhibition of renal human hyaluronidase-1 by isoflavonoids. Preliminary studies also indicated anti-inflammatory effects. The following study aimed at investigating potential anti-inflammatory effects of restharrow extracts, prepared with solvents of different polarity. A dichloromethane extract (OS1), mainly composed of isoflavonoids and triterpenes as characterized by LC-MS, showed a concentration-dependent (25–100 μg/ml) inhibition of IL-8 and TNF-α release from LPS-stimulated human neutrophils. Significant inhibition was also found for the triterpene α-onocerin and the norneolignan clitorienolactone B, isolated from OS1. Further, OS1 and both compounds significantly decreased the expression of the adhesion molecules CD11b/CD18 and conversely increased the expression of CD62L in LPS-stimulated human neutrophils. This finding corresponds to a reduced inflammatory response by the inhibition of adhesion and migration of immune cells. As all of the observed effects are potentially mediated via Toll-like receptor 4 (TLR4) signaling, TLR4 transfected HEK293 cells were incubated with OS1. LPS-induced IL-8 secretion was significantly inhibited in a concentration-dependent manner, confirming TLR4 antagonism. This inhibition, however, was in part caused by an interaction of OS1 with LPS. In addition, also an aqueous extract containing high amounts of isoflavonoid glycosides and saponins from the roots of O. spinosa showed anti-inflammatory effects by interacting with the TLR4 signaling pathway. This study rationalizes the traditional use of extracts from O. spinosa for therapy of urinary tract infections, due to its potential anti-inflammatory effects that are mediated via TLR4 receptor antagonism.

This study was to obtain stable transformed roots of Salvia bulleyana using A. rhizogenes strain A4 and then evaluate their phytochemical profile and selected the most productive clone. Our results indicated that the type of explant and medium used for bacterium and explant incubation had an influence on the frequency of hairy root formation. The best response was obtained on leaves infected with bacteria cultivated on YMB medium supplemented with acetosyringone. Of the four selected transformed root clones, after five-week cultivation in Woody Plant (WP) medium, the highest growth indexes were demonstrated for line C1: i.e. 13 for fresh and 15 for dry weight (81.4 and 8.2 g/l fresh and dry weight, respectively). The qualitative analysis of hydromethanolic extracts of hairy roots of S. bulleyana using UPLC-PDA-ESI-MS/MS method showed the presence of 10 polyphenolic compounds including predominant rosmarinic acid (RA), its derivatives (hexoside and methyl rosmarinate), caffeic acid, its derivatives and several salvianolic acids: K, E and F. Their production varied among the four root clones studied; the highest RA (39.6 mg/g dry weight) and total polyphenol (48.9 mg/g dry weight) level were found in the roots of C4 clone. These values were significantly higher than those of the roots of plants grown for several years under field conditions. The transformation of the obtained root cultures was confirmed by polymerase chain reaction using aux1, aux2, rolB, rolC and rolD primers.

Acmella oleracea (jambu) is a South American plant whose pharmacological properties include antimalarial, anesthetic, and anti-inflammatory activities. However, the compounds contributing to the anti-inflammatory effect have received little attention. In this study, the active compound spilanthol was isolated using a Green Chemistry approach and presented significant anti-inflammatory activity in a lipopolysaccharide-activated human neutrophil model. First, the aerial parts of jambu were extracted and fractionated using supercritical carbon dioxide. The spilanthol-enriched fractions obtained were then subjected to further purification by flash chromatography, yielding spilanthol with 97% purity. Use of enzyme-linked immunosorbent assay (ELISA) revealed a reduction in the release of Interleukin-8 and Tumor Necrosis Factor - alpha by leukocytes exposed to spilanthol. In conclusion, this new approach enabled spilanthol to be obtained at high purity, in a fairly rapid procedure, while the results of the in vitro anti-inflammatory activity study indicated that the compound could be a promising new therapeutic agent.

Thirteen compounds were isolated and identified from the aerial parts of Eupatorium cannabinum subsp. cannabinum comprising six caffeic acid derivatives, five flavonol glycosides and two flavone aglycones. Additionally, the first HPLC-DAD-MS method for the phytochemical screening of phenolics in the analyzed plant material was developed allowing unequivocal identification of further six constituents occurring in the raw extract including four caffeic acid derivatives and two malonylated flavonol glycosides. This report is the first comprehensive study focused on phenolic compounds in the aerial parts of E. cannabinum. The chemotaxonomic significance in respect to the genus Eupatorium is discussed.

Salvia viridis is a containing bioactive phenols plant, which has been used in Turkish medicine. The aim of the study was to establish hairy root culture of the species in order to increase the production of the compounds with pharmacological activity. Transformed roots are promising biotechnological systems that produce great amounts of secondary metabolites; this is attributed to their fast growth and their genetic and biosynthetic stability. Stable hairy root lines of S. viridis were established from shoot explants using Agrobacterium rhizogenes strain A4. The highest frequency of transformation (45%) was achieved when the shoot was inoculated at the node or internode by bacteria cultivated on Yeast/Mannitol/Broth (YMB) medium supplemented with acetosyringone. The transformation of the root clones was confirmed by polymerase chain reaction using aux1, aux2, rolB and rolC primers. Of the five obtained transformed root clones, the highest biomass was achieved for clone K3 (13.63 ± 0.5 g/L after 5 weeks) grown in Woody Plant (WP) medium in darkness. The UPLC-PDA-ESI–MS analysis of hydromethanolic extracts of transformed roots of S. viridis revealed the presence of 10 compounds identified as caffeic acid derivatives. Quantitative analysis showed that rosmarinic acid (RA) was the predominant compound in all clones. The highest RA (35.8 mg/g dry weight) and total polyphenol (41.24 mg/g dry weight) content were evaluated in clone K3 cultured in WP medium. These values were 8-fold higher than those of the non-transformed roots, indicating that hairy root cultivation could be a promising technique for the production of valuable compounds from S. viridis.

Epilobium sp. are commonly used in traditional medicine in the treatment of early stages of benign prostatic hyperplasia and inflammation. It is suggested that a dominating constituent, oenothein B, is responsible for the extracts therapeutic effects. Several bioactivities were established for extracts and oenothein B in various in vitro models, but due to the questionable bioavailability of this dimeric macrocyclic ellagitannin, their significance in the in vivo effects remains unresolved. We have thus focused our attention on a complex comparative investigation of the in vitro and in vivo activities of phytochemically characterized Epilobium angustifolium aqueous extract and oenothein B on prostate cancer cells proliferation.Incubation of different cell lines with E. angustifolium aqueous extract resulted in a significant reduction of proliferation of PZ-HPV-7 and LNCaP cells, which was partly associated with antiandrogenic activity. These effects were fully congruent with oenothein B, examined in parallel. Oral supplementation of rats implanted with LNCaP cells with E. angustifolium aqueous extract 50–200 mg/kg b. w. resulted in a reduction of the occurrence of prostatic adenoma up to 13 %. Oenothein B was not detected in the urine and feces of the E. angustifolium aqueous extract-treated group, however, conjugates of nasutins gut microbiota metabolites of ellagitannins were detected in the urine, while in human volunteers supplemented with Epilobium tea, only urolithin conjugates were present.Despite observing significant and consistent effects in vitro and in vivo, we were unable to point out unequivocally the factors contributing to the observed E. angustifolium aqueous extract activity, facing the problems of an unknown metabolic fate of oenothein B and interspecies differences in E. angustifolium aqueous extract gut microbiota metabolism.

Plants belonging to the family Oleaceae have been traditionally used in treatment of various inflammatory skin disorders. One of the most well-known species is Olea europaea L. (olive), cultivated in the Mediterranean countries. Another species is Ligustrum vulgare L. (common privet), occurring particularly in Northern Europe and Asia. The aim of the study was a comparison of the composition of aqueous and ethanolic extracts from leaves of O. europaea and L. vulgare (HPLC-DAD-MS), and determination of the total content of phenolics and flavonoids, as well as the content of the major compound, oleuropein. Secondly, we aimed to study the protective effect of extracts on reactive oxygen species (ROS) production by human fibroblast cells (NHDF), cell viability (MTT assay), and apoptosis rate (Annexin V/propidium iodide staining) after UVA-irradiation. The phytochemical analysis allowed us to identify compounds belonging to the groups of flavonoids, phenylpropanoids and secoiridoids in the extracts. The compounds from the group of lignans (olivil) were identified as being unique to O. europaea extracts. Echinacoside, ligustroflavone and ligustaloside A were identified in L. vulgare extracts in contrast to olive preparations. It was established that the aqueous and ethanolic extracts from leaves of both species, except the privet aqueous extract at a concentration of 5 μg/ml, did not show any significant inhibition of ROS production after UVA-irradiation in the model of NHDF cell line. The aqueous extracts of both species at concentrations of 5 and 25 μg/ml had a protective effect on the viability of UVA-treated cells in contrast to the ethanolic extract. In conclusion, no significant difference in the activity of olive and privet leaf extracts has been observed, which suggests that both plant materials’ extracts, particularly aqueous ones, are effective herbal medicines and photoprotectors, which – to some extent – confirms the use of their preparations in skin disorders.

Ethnopharmacological relevance During the epidemic season, over 90% of acute wheezing disease is associated with bronchial inflammation. Both neutrophil- and eosinophil-mediated inflammation have been involved in the pathophysiology of acute bronchitis, but neutrophil cell recruitment has been shown to be dominant. The ongoing inflammation increases the chemotaxis of neutrophils to inflamed site providing to their overaccumulation. The pharmacological reduction of neutrophil migration can be limited by suppression of major chemo-attractants and cytokines (IL-8, IL-1β and TNF-α) release and downregulation of adhesive molecules. Aim of the study During a screening of plants traditionally used in respiratory tracts diseases (e.g. cough, rhinitis, bronchitis, throat infection, fever, influenza) in Europe, we have selected roots of Inula helenium and aerial parts of Grindelia squarrosa as a potential source of compounds limiting neutrophil migration. Materials and methods The effect on IL-8, IL-1β and TNF-α release by neutrophils and respiratory epithelium cell line (A549) was measured by enzyme-linked immunosorbent assay (ELISA). The surface expression of adhesive molecules was analyzed with flow cytometry, and the neutrophil attachment to the epithelial cells was assessed fluorimetrically. Results We confirmed the ability of selected extracts and compounds to suppress neutrophil binding to the epithelium surface via downregulation of β2 integrin. Alantolactone and grindelic acid have shown significant suppression of IL-8, TNF-α and IL-1β release comparable with budesonide, used as a positive control. Conclusions The present study demonstrated that Inula helenium and Grindelia squarrosa, which have been traditionally used in Europe as medicinal plants, are a valuable source of active compounds with anti-inflammatory activity. Our observations justify the traditional use of I. helenium and G. squarrosa for a treatment of inflammation-based diseases in respiratory tract.

During chronic inflammation, neutrophils acting locally as effector cells not only activate antibacterial defense but also promote the inflammatory response. Interleukin 8 (IL-8), the main cytokine produced by activated neutrophils, positively correlates with the severity of respiratory tract diseases. By screening European plants traditionally used for treating respiratory tract diseases, we found that extracts of aerial parts of Eupatorium cannabinum inhibit IL-8 release from neutrophils. Using bioassay-guided fractionation, we identified five sesquiterpene lactones, eupatoriopicrin (1), 5′-deoxyeupatoriopicrin (2), hiyodorilactone A (3), 3-hydroxy-5′-O-acetyleupatoriopicrin = hiyodorilactone D (4), and hiyodorilactone B (5), that efficiently (IC50 < 1 μM) inhibited IL-8 and TNF-α release in lipopolysaccharide (LPS)-stimulated human neutrophils. Moreover, all these sesquiterpene lactones suppressed the adhesion of human neutrophils to an endothelial monolayer by downregulating the expression of the β2 i...

Background and purpose Hypertension is a multifactorial disease, manifested by vascular dysfunction, increased superoxide production, and perivascular inflammation. In this study, we have hypothesized that 1,2,3,4,6-penta-O-galloyl-β-d-glucose (PGG) would inhibit vascular inflammation and protect from vascular dysfunction in an experimental model of hypertension. Experimental approach PGG was administered to mice every 2 days at a dose of 10 mg·kg-1 i.p during 14 days of Ang II infusion. It was used at a final concentration of 20 μM for in vitro studies in cultured cells. Key results Ang II administration increased leukocyte and T-cell content in perivascular adipose tissue (pVAT), and administration of PGG significantly decreased total leukocyte and T-cell infiltration in pVAT. This effect was observed in relation to all T-cell subsets. PGG also decreased the content of T-cells bearing CD25, CCR5, and CD44 receptors and the expression of both monocyte chemoattractant protein 1 (CCL2) in aorta and RANTES (CCL5) in pVAT. PGG administration decreased the content of TNF+ and IFN-γ+ CD8 T-cells and IL-17A+ CD4+ and CD3+ CD4- CD8- cells. Importantly, these effects of PGG were associated with improved vascular function and decreased ROS production in the aortas of Ang II-infused animals independently of the BP increase. Mechanistically, PGG (20 μM) directly inhibited CD25 and CCR5 expression in cultured T-cells. It also decreased the content of IFN-γ+ CD8+ and CD3+ CD4- CD8- cells and IL-17A+ CD3+ CD4- CD8- cells. Conclusion and implication PGG may constitute an interesting immunomodulating strategy in the regulation of vascular dysfunction and hypertension. Linked articles This article is part of a themed section on Immune Targets in Hypertension. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v176.12/issuetoc.

Natural phenolic compounds exhibit a lot of beneficial health effects in humans including antioxidant, anti-aging, anti-inflammatory, and antimicrobial activity. Plant family Lamiaceae and its largest genus Salvia are one of the richest sources of antioxidant and antimicrobial phenolics. In the present study the aerial parts and roots of Salvia cadmica Boiss., an underexplored species endemic to Turkey and cultivated in Poland, were investigated for phenolic composition and antioxidant, antibacterial, and antifungal effects, as well as for biological safety. The UPLC-DAD/ESI-MS/MS analysis showed a total of 14 phenols in the hydromethanolic extract of the aerial parts: three flavonoids and 11 phenolic acid derivatives. In the root extract 10 phenolic acid derivatives were identified. Among the detected compounds, rosmarinic acid (RA) and salvianolic acid K (SAK) predominated in the aerial parts and roots, respectively. Both aerial parts and root extracts demonstrated antioxidant potential, indicated by radical scavenging assays towards DPPH and O2•-; they also demonstrated antimicrobial activity against two Candida species and several Gram-positive bacteria, including Bacillus cereus and four strains of Staphylococcus spp. However, the aerial parts demonstrated stronger antioxidant potential, while the roots exhibited stronger antimicrobial activity The tested extracts show low cytotoxicity against mouse L929 fibroblasts in MTT assay. Thus, S. cadmica extracts offer promise as herbal drug, and as a source of bioactive secondary metabolites.

Dracocephalum forrestii, an important plant in Tibetan medicine, was transformed with Agrobacterium rhizogenes strain A4 to obtain hairy root culture. A maximum transformation frequency of 82% after seven weeks was obtained by inoculation of internodal stem segments. Six lines of hairy roots were obtained and cultured in darkness in Woody Plant (WPM) liquid medium. The transgenic nature of the hairy roots was investigated by PCR and RT-PCR. Three main compounds: rosmarinic acid (RA), salvianolic acid B (SAB) and caffeic acid (CA) were identified in extracts of D. forrestii hairy roots by UPLC-PDA-ESI-MS analysis. Quantitative HPLC analysis showed that RA levels were greatly increased in hairy roots (max. 19.97 mg/g DW) compared with the roots of one-year-old field grown plants (4.7 mg/g DW), but SAB levels were significantly lower in transformed roots (max. 11.7 mg/g DW) than in untransformed roots (16.4 mg/g DW). Caffeic acid was detected only in transformed roots. A time course experiment with the most productive hairy root line showed that a culture period of 30 days and a medium supplemented with 3% of sucrose was optimal for biomass and production of analysed phenolic compounds. Production remained stable over at least 3.5 years of continuous subculturing of the hairy roots.