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The hawthorn ( Crataegus spp.) extract WS 1442 is used against mild forms of chronic heart failure. This disease is associated with endothelial barrier dysfunction and edema formation. We have recently shown that WS 1442 protects against this dysfunction by a dual mechanism: it both promotes endothelial barrier integrity by activation of a barrier-enhancing pathway (cortactin activation) and inhibits endothelial hyperpermeability by blocking a barrier disruptive pathway (calcium signaling). In this study, we aimed to identify the bioactive compounds responsible for these actions by using a bioactivity-guided fractionation approach. From the four fractions generated from WS 1442 by successive elution with water, 95 % ethanol, methanol, and 70 % acetone, only the water fraction was inactive, whereas the other three triggered a reduction of endothelial hyperpermeability. Analyses of intracellular calcium levels and cortactin phosphorylation were used as readouts to estimate the bioactivity of subfractions and isolated compounds. Interestingly, only the ethanolic fraction interfered with the calcium signaling, whereas only the methanolic fraction led to an activation of cortactin. Thus, the dual mode of action of WS 1442 could be clearly assigned to two distinct fractions. Although the identification of the calcium-active substance(s) was not successful, we could exclude an involvement of phenolic compounds. Cortactin activation, however, could be clearly attributed to oligomeric procyanidins with a distinct degree of polymerization. Taken together, our study provides the first approach to identify the active constituents of WS 1442 that address different cellular pathways leading to the inhibition of endothelial barrier dysfunction., (Georg Thieme Verlag KG Stuttgart · New York.)

Porphyromonas gingivalis is a pathogen strongly involved in chronic and aggressive forms of periodontitis. Natural products, mainly polyphenols, have been described for advanced treatment of periodontitis by inhibition of the bacterial adhesion of P. gingivalis to the epithelial host cells. An acetone:water extract (LBE) from the rhizomes of Limonium brasiliense (Boiss.) Kuntze was tested under in vitro conditions for potential antiadhesive effects against P. gingivalis to human KB cells and for inhibition of the proteolytic activity of gingipains, the main virulence factor of P. gingivalis. LBE≤100μg/mL had no cytotoxicity against the bacteria and did not influence the cell physiology of human epithelial KB cells. At 100μg/mL LBE reduced the adhesion of P. gingivalis to KB cells significantly by about 80%. LBE at 20μg/mL reduced the proteolytic activity of the arginin-specific Rgp gingipain by about 75%. Chemical profiling of LBE indicated the presence of gallic acid, epigallocatechin-3-O-gallate and samarangenins A and B as lead compounds. UHPLC by using MS and UV detection displays a suitable method for quality control of the extract for identification and quantification of the lead compounds., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Extract RE was obtained from the roots of Althaea officinalis in a yield of 8.1%, related to the dried plant material, by extraction with MeOH-H 2 O (1:1), followed by precipitation with EtOH to remove high molecular weight constituents. Phytochemical investigation of RE revealed the presence of N-phenylpropenoyl-l-amino acid amides 1-5, 8% glycine betaine 6, about 9% total amino acids with proline as the main compound, and about 61% mono- and oligomeric carbohydrates with sucrose as the main compound. Further fractionation revealed the presence of a hypolaetin diglycoside (12) and four hypolaetin glycosides (7-9 and 11) with O-sulfocarbohydrate moieties; additionally, 4'-O-methylisoscutellarein-8-O-β-d-(3″-O-sulfo)glucuronopyranoside (10) and the diglycosylated coumarin haploperoside D (13) were identified. The hypolaetin-O-sulfoglycosides 7-10 are new natural products. RE inhibited the enzymatic activity of surface-displayed human hyaluronidase-1 on Escherichia coli F470 cells with an IC 50 of 7.7 mg/mL. RE downregulated mRNA expression of hyal-1 in HaCaT keratinocytes at 125 and 250 μg/mL, respectively. These data contribute to a deeper phytochemical understanding of marshmallow root extracts and to the positive influence of extracts used for therapy of irritated and inflamed buccal tissue and cough.

The upper aerial parts and leaf tips of Myrothamnus flabellifolia Welw. (Myrothamnaceae), a resurrection plant indigenous to southern Africa, are used in African traditional medicine for infections of the respiratory and urinary system as well as for inflammation of mucosa and skin. Within a phytochemical investigation of the herbal material from M. flabellifolia the flavonoid fraction was shown to contain quercetin 10 as well as the respective 3-O-β-d-galactosides, glucosides, -glucuronides and 3-O-α-l-rhamnosides of quercetin (6, 7, 8, 26) and kaempferol (1, 2, 3, 9). Additionally mono-galloylated 12, 13, di-galloylated 15 and tri-galloylated 16 flavonol glycosides were identified. 3,4,5-Tri-O-galloyl- 14 and 1,3,4,5-tetra-O-galloyl quinic acid 28 were isolated and characterized, beside arbutin 27 and 2″,3″-di-O-galloyl-arbutin 11. Furthermore, the depside 2-O-(3,4-dihydroxybenzoyl)-2,4,6-trihydroxyphenylacetic acid 5, 1,2,3,4,6-penta-O-galloyl-β-d-glucose 22 and seven ellagitannins were found: 1,2-di-O-galloyl-(4,6-(S)-hexahydroxydiphenoyl)-β-d-glucose 20, 1,3-di-O-galloyl-(4,6-(S)-hexahydroxydiphenoyl)-β-d-glucose 21, 2,3-di-O-galloyl-(4,6-(S)-hexahydroxydiphenoyl)-β-d-glucose (tellimagrandin-I) 19, 1,2,3-tri-O-galloyl-(4,6-O-hexahydroxydiphenoyl)-β-d-glucose (tellimagrandin-II) 23, and two so far not described dimers of tellimagrandin-I and -II (myrodigamin-I and -II, 24, 25). The presence of trehalose (3.3%), raffinose (0.2%) and stachiose (0.2%) beside a fructan (2.1%) was determined. Two ICH2-validated UHPLC methods have been developed and used within batch analysis for unambiguous identification of the herbal material and quantification of the major compounds myrodigamin-I (0.9 to 1.7%), tellimagrandin-II (0.3 to, 0.9%) and 3,4,5-tri-O-galloyl quinic acid (0.1 to 0.7%), besides kaempferol (0.1 to 0.3%) and quercetin content (0.2 to 0.3%) after hydrolysis of the respective glycosides., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Olax mannii Oliv. (Olacaceae) is among the many medicinal plants used in Nigeria for the ethnomedicinal management of both cancer and inflammation. Such plants represent potential sources of innovative therapeutic agents for the treatment of cancer and other malignant disorders. While the majority of medicinal plants exert their anticancer effects by direct cytotoxicity on tumor cells, it is important that other mechanisms through which these plants can exhibit anticancer effects are investigated. Preliminary studies indicated that Olax mannii leaves are rich sources of novel flavonoid glycosides. The detailed chemistry as well the mechanisms through which these flavonoid constituents may exert their cancer chemo-preventive and therapeutic effects are, however, not yet investigated., Aim of the Study: The aim of this study is to carry out a detailed chemical investigation of Olax mannii leaves and the effects of the isolated constituents on the nuclear factor kappa B (NF-κB) pathway., Materials and Methods: A methanol leaf extract was subjected to various chromatographic separations to achieve isolation of flavonoid glycosides and the structures of the isolated compounds were elucidated by a combination of 1D and 2D NMR and high resolution mass spectrometry. Biological activities were assessed by measurement of cellular viability and proliferation using quantitative IncuCyte videomicroscopy, trypan blue staining and by quantification of the number of metabolically active K562 cells based on quantitation of ATP. The effect of the compounds on the inhibition of the NF-κB pathway as well as toxicity towards peripheral blood mononuclear cells to evaluate differential toxicity was also assayed., Results: Chemical investigation of the methanol leaf extract of the plant material led to the isolation of three new flavonoid triglycosides, kaempferol 3-O-[α-D-apiofuranosyl-(1 → 2)-α-L-arabinofuranoside]-7-O-α-L-rhamnopyranoside (1), kaempferol 3-O-[β-D-glucopyranosyl-(1 → 2)-α-L-arabinofuranoside]-7-O-α-L-rhamnopyranoside (2), kaempferol 3-O-[β-D-arabinopyranosyl-(1→4)-α-L-rhamnopyranoside]-7-O-α-L-rhamnopyranoside (3), in addition to fourteen known flavonoid glycosides (4-17). Of all the tested compounds, only compound 9 (kaempferol 3-O-α-L-rhamnopyranoside) exhibited promising and specific antiproliferative activity on human K562 chronic myelogenous leukemia cells and dose-dependently inhibited NF-κB transactivation., Conclusion: The presence of this flavonoid glycoside and derivatives may account for the reported efficacy of Olax mannii leaf extract in the ethnomedicinal management of cancer and inflammation., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Leaves from Phyllanthus muellerianus (Kuntze) Exell. are traditionally used for wound healing in Western Africa. Aqueous extracts of dried leaves recently have been shown to stimulate proliferation of human keratinocytes and dermal fibroblasts. Within bioassay-guided fractionation the ellagitannins geraniin (1), corilagin (2), furosin (3), the flavonoids quercetin-3-O-β-D-glucoside (isoquercitrin), kaempferol-3-O-β-D-glucoside (astragalin), quercetin-3-O-D-rutinoside (rutin), gallic acid, methyl gallate, caffeic acid, chlorogenic acid, 3,5-dicaffeoylquinic acid and caffeoylmalic acid (phaselic acid) have been identified in P. muellerianus for the first time. Geraniin was shown to be the dominant component of an aqueous extract. Suitable analytical methods for quality control of geraniin in P. muellerianus extract (methanol/water, 70/30) have been developed and validated based on ICH guidelines (ICH-compliant protocol). Geraniin and furosin increased the cellular energy status of human skin cells (dermal fibroblasts NHDF, HaCaT keratinocytes), triggering the cells towards higher proliferation rates, with fibroblasts being more sensitive than keratinocytes. Highest stimulation of NHDF by geraniin was found at 5 μM, and of keratinocytes at 50-100 μM. Furosin stimulated NHDF at about 50 μM, keratinocytes at about 150-200 μM. Necrotic cytotoxicity of geraniin, as measured by LDH release, was observed at 20 μM for NHDF and 150 μM for keratinocytes. Toxicity of furosin--less than that of geraniin--was observed at > 400 μM. Furosin and geraniin stimulated the biosynthesis of collagen from NHDF at 50 μM and 5-10 μM respectively. Geraniin at 105 μM significantly stimulated the differentiation in NHEK while furosin had a minor influence on the expression of involucrin and cytokeratins K1 and K10. The study proves clearly that hydrophilic extracts from P. muellerianus and especially the lead compound geraniin exhibit stimulating activity on dermal fibroblasts and keratinocytes, leading to increased cell proliferation, barrier formation and formation of extracellular matrix proteins. From these findings the traditional clinical use of such extracts for wound healing seems to be justified., (Copyright © 2010 Elsevier GmbH. All rights reserved.)

Aim: Identification of anti-adhesive plant extracts against cell surface binding of Porphyromonas gingivalis and underlying mechanisms; investigation of potential cytoprotective effects of anti-adhesive extract on KB cells., Materials and Methods: Polyphenol-enriched extract, fully characterized concerning flavan-3-ols and oligomeric proanthocyanidins, from Myrothamnus flabellifolia (MF), traditionally used for periodontitis, was tested for inhibition of P. gingivalis-mediated adhesion to KB cells by flow cytometry, for influence on gingipain activity (protease assay), haemagglutination and by microarray analysis for effects on bacterial transcriptome. The influence of MF on P. gingivalis-induced cytokine gene expression was monitored by RT-PCR and IL-6 titres by ELISA., Results: MF (100 μg/ml) reduced P. gingivalis adhesion/invasion about 50% by interacting with bacterial OMPs. As shown by RT-PCR, fimbrillin and Arg-gingipain encoding genes were up-regulated by MF. On the protein level, inhibition (70%) of Arg-gingipain activity was observed, while the corresponding Lys-gingipain was hardly influenced. MF also inhibited haemagglutination. While exposure to P. gingivalis resulted in an increased expression of inflammation-related genes in KB cells, pre-treatment of KB cells with MF evoked cytoprotective effects concerning IL-1β, IL-6, IL-8 and TNF-α gene expression as well as IL-6 release rates. Compounds from the plant extract belonging to the class of proanthocyanidins were shown to be responsible for the observed effects and were characterized for their respective structural features., Conclusions: While being cytoprotective, MF exerts anti-adhesive effects against P. gingivalis. Thus, MF may be useful for the prevention of P. gingivalis-associated periodontal diseases., (© 2010 John Wiley & Sons A/S.)

Within a systematic phytochemical investigation of the leaves of RHODODENDRON FERRUGINEUM L. (Ericaceae), the volatile oil was isolated (0.7 %) and its constituents were characterized. Eleven flavonoids were isolated and characterized, with quercetin 3- O-[6''- O-(2-methyl-1-oxobutyl)]-β- D-glucopyranoside and 2 R,3 R-dihydromyricetin 3- O- β- L-arabinopyranoside as new natural products. Beside monomeric flavan-3-ols (catechin, epicatechin, gallocatechin, epigallocatechin) from the tannin fraction (about 3.5 % calculated as pyrogallol), the dimeric procyanidins B1 to B7 were identified, as well as the trimeric compounds procyanidin C1, epicatechin-(4 β → 8)-epicatechin-(4 β → 8)-catechin and the trimeric A type-linked cinnamtannin B1. Additionally, phloroacetophenon 4- O- β- D-glucopyranoside and chlorogenic acid were isolated. Water-soluble carbohydrates comprised about 13.5 % of the dried leaves, including fructans (3 %), polysaccharides (1 %) (mainly type II arabinogalactans), glucose, fructose, sucrose, stachyose and raffinose. The IN VITRO effects on cellular vitality (MTT test), proliferation (BrdU incorporation) and necrosis (LDH release) of an aqueous extract were investigated. The extract did not exert any toxic effects, while the vitality and the proliferation rates of epithelial HaCaT keratinocytes were significantly increased at 100 µg/mL, indicating that the aqueous extract does not have negative effects against cellular activity., (© Georg Thieme Verlag KG Stuttgart · New York.)

Besides flavan-3-ols, a family of N-phenylpropenoyl-L-amino acids (NPAs) has been recently identified as polyphenol/amino acid conjugates in the seeds of Theobroma cacao as well as in a variety of herbal drugs. Stimulated by reports on their biological activity, the purpose of this study was to investigate if these amides are absorbed by healthy volunteers after administration of a cocoa drink. For the first time, 12 NPAs were quantified in human urine by means of a stable isotope dilution analysis with LC-MS/MS (MRM) detection. A maximum amount was found in the urine taken 2 h after the cocoa consumption. The highest absolute amount of NPAs excreted with the urine was found for N-[4'-hydroxy-(E)-cinnamoyl]-L-aspartic acid (5), but the highest recovery rate (57.3 and 22.8%), that means the percentage amount of ingested amides excreted with the urine, were determined for N-[4'-hydroxy-(E)-cinnamoyl]-L-glutamic acid (6) and N-[4'-hydroxy-3'-methoxy-(E)-cinnamoyl]-L-tyrosine (13). In order to gain first insights into the NPA metabolism in vivo, urine samples were analyzed by LC-MS/MS before and after beta-glucuronidase/sulfatase treatment. As independent of the enzyme treatment the same NPA amounts were found in urine, there is strong evidence that these amides are metabolized neither via their O-glucuronides nor their O-sulfates. In order to screen for caffeic acid O-glucuronides as potential NPA metabolites, urine samples were screened by means of LC-MS/MS for caffeic acid 3-O-beta-D-glucuronide and 4-O-beta-D-glucuronide. But not even trace amounts of one of these glucuronides were detectable, thus excluding them as major NPA metabolites and underlining the importance of future investigations on a potential O-methylation or reduction of the N-phenylpropenoyl moiety in NPAs.

Extracts from Drosera rotundifolia are traditionally used to treat cough symptoms during a common cold. The present study aimed to investigate the impact of extracts from D. rotundifolia and active compounds on the respiratory tract. Tracheal slices of C57BL/6N mice were used ex vivo to examine effects on airway smooth muscle (ASM) and ciliary beat frequency (CBF). Phosphodiesterase (PDE) inhibition assays were carried out to test whether PDE1 or PDE4 are targeted by the active compounds. An ethanol–water extract, as well as an aqueous fraction of this extract, exerted antispasmodic properties against acetylcholine-induced contractions. In addition, contractions induced by 60 mM K+ were abrogated by the aqueous fraction. Effects on ASM could be attributed to the flavonoids quercetin, 2″-O-galloylhyperoside and hyperoside. Moreover, the Drosera extract and the aqueous fraction increased the CBF of murine tracheal slices. Quercetin and 2″-O-galloylhyperoside were identified as active compounds involved in the elevation of CBF. Both compounds inhibited PDE1A and PDE4D. The elevation of CBF was mimicked by the subtype-selective PDE inhibitor rolipram (PDE4) and by 8-methoxymethyl-IBMX. In summary, our study shows, for the first time, that a Drosera extract and its flavonoid compounds increase the CBF of murine airways while antispasmodic effects were transferred to ASM. [ABSTRACT FROM AUTHOR]

Combretum mucronatum Schumach. & Thonn. is a medicinal plant widely used in West African traditional medicine for wound healing and the treatment of helminth infections. The present study aimed at a phytochemical characterization of a hydroalcoholic leaf extract of this plant and the identification of the anthelmintic compounds by bioassay-guided fractionation. An EtOH-H2O (1:1) extract from defatted leaves was partitioned between EtOAc and H2O. Further fractionation was performed by fast centrifugal partition chromatography, RP18-MPLC and HPLC. Epicatechin (1), oligomeric proanthocyanidins (OPC) 2 to 10 (mainly procyanidins) and flavonoids 11 to 13 were identified as main components of the extract. The hydroalcoholic extract, fractions and purified compounds were tested in vitro for their anthelmintic activity using the model nematode Caenorhabditis elegans. The bioassay-guided fractionation led to the identification of OPCs as the active compounds with a dose-dependent anthelmintic activity ranging from 1 to 1000 μM. Using OPC-clusters with a defined degree of polymerization (DP) revealed that a DP ≥ 3 is necessary for an anthelmintic activity, whereas a DP > 4 does not lead to a further increased inhibitory effect against the helminths. In summary, the findings rationalize the traditional use of C. mucronatum and provide further insight into the anthelmintic activity of condensed tannins. [ABSTRACT FROM AUTHOR]

Abstract: Ethnopharmacological relevance: Eupatorium perfoliatum L. originates from North America, where it has been widely used since centuries by native Indians. Additionally extracts are used also in Europe as immunostimulating agent for treatment of fever and cold. The following review summarizes published data on phytochemistry, ethnopharmacological use, as well as clinical and preclinical data. Materials and methods: Literature survey was performed via SciFinder® on papers and patents and by systematic research in ethnopharmacological literature at various university libraries. Results: The phytochemical composition of Eupatorium perfoliatum is described in detail for volatile oil, caffeic acid derivatives, flavonoids, sesquiterpene lactones, tannins, polysaccharides. Methods for analytical quality control, as well as specification for relevant lead structures can be deduced from published batch analysis. Preclinical studies indicate anti-inflammatory effects of ethanolic extracts, which can be correlated on a molecular level to eupafolin and sesquiterpen lactones. Antiplasmodial, antioxidative and immunomodulating activities are additionally discussed. Clinical data on the use of Eupatorium perfoliatum do not meet modern GCP requirements, but do indicate positive tendencies for use of ethanolic extracts for treatment of common colds. Conclusion: While the postulated immunostimulating properties of Eupatorium perfoliatum have not been confirmed by in vitro data, animal-studies and in vitro experiments with plant extracts both indicate antiinflammatory effects beside antiplasmodial effect against Plasmodium falciparum. Such an antiinflammation caused by the ethanolic extracts can be correlated well with clinical symptoms related to diseases as common cold, rheumatism, athritis etc. These data also support the plausibility of the plant''s traditional use by the North American indigenous population and early European settlers. In principle quality aspects of the herbal material have to be affirmed by establishing modern pharmacopoeial control methods to guarantee constant and reliable quality. [Copyright &y& Elsevier]

Abstract: The CH2Cl2 extract of aerial parts of Eupatorium perfoliatum L. exhibits antiprotozoal activity under in vitro conditions, especially against Plasmodium falciparum (IC50 =2.7μg/ml). The search for active compounds yielded seven sesquiterpene lactones: Four structurally similar guaianolides, one dimeric guaianolide, and two germacranolides. The guaianolides differ in the degree of oxidation at C-14, ranging from a hydroxyl group up to a free carboxylic acid. The dimeric guaianolide, structurally closely related to the monomers, displays an unusual type of interguaianolide linkage between C-14 and C-4. Except for the germacranolide euperfolitin, all STLs described here were hitherto unknown. Furthermore, the flavonoid aglycones eupafolin, hispidulin, patuletin, and kaempferol were identified in the extract, which, except for kaempferol, have not been described as constituents of E. perfoliatum before. The dimeric guaianolide was shown to be the most active constituent against Plasmodium falciparum (IC50 = 2.0μM) and was less cytotoxic against rat skeletal myoblasts (IC50 = 16.2μM, selectivity index of about 8). [Copyright &y& Elsevier]

Ethnopharmacological relevance The aerial parts of Lippia integrifolia (Gris.) Hieronymus (Verbenaceae), known as incayuyo , are used by the peasant population of Argentina for treatment of diseases related to a gastrointestinal system, mainly for inflammation of the stomach and have also been included into the Argentina Food Code. This study aimed to investigate the phytochemical profile of hydrophilic extracts from the herbal material by LC−MS and to evaluate potential pharmacological mechanisms rationalizing the traditional use of incayuyo aqueous extracts potential anti-inflammatory treatment of gastrointestinal disorders. Materials and methods Phytochemical profiling: LC−MS of an aqueous decoction. Antiadhesive effects against Helicobacter pylori : in vitro FACS assay using FITC-labeled bacteria and AGS human stomach cells. Influence of extracts on stomach cells and RAW 264.7 macrophages: MTT viability assay and BrdU proliferation ELISA. Influence of extracts on IL-6 and IL-8 secretion from stomach cells was quantitated by ELISA after infection of the cells with Helicobacter pylori . Influence of test extracts on macrophages: phagocytosis of FITC-labeled Zymosan particles and NO production. Antioxidative capacity: DPPH assay and O 2 -induced caroten oxidation. Results LC−MS profiling indicated the presence of compounds 1 − 20 with flavonoid hexosides, phenylethanoides (acteoside, isoacteoside) and sesquiterpenes [(epi)lippidulcine, peroxylippidulcine] in the decoction extract and subfraction PhF. The extract exhibits strong in vitro antioxidative capacity and inhibited adhesion of Helicobacter pylori to stomach cells up to 40%, while an EtOH-soluble fraction showed inhibition rates of up to 60%. Decoction increased the cellular viability of AGS cells significantly at >10 μg/mL, while proliferation rate was not influenced. Helicobacter pylori induced IL-8 secretion was significantly reduced by coincubation of AGS cells with the extracts. Aqueous extracts stimulated phagocytosis rate of macrophages and inhibited the LPS-induced NO-secretion. Conclusions The traditional use of aqueous extracts from Lippia integrifolia for gastric inflammation seems to be rationalized: besides anti-inflammatory effects on stomach cells antiadhesive properties of the extracts against the main bacterial inductor of gastritis Helicobacter pylori were identified. Additional effects for stimulation of innate immunity and potential radical scavenging effects may additionally contribute to the activity of the extracts. [ABSTRACT FROM AUTHOR]