Antifibrotic activity of triterpenoids from the aerial parts of Euscaphis japonica on hepatic stellate cells

In the course of screening antifibrotic activity of natural products employing HSC-T6, a rat hepatic stellate cell line, as an in vitro assay system, the methanolic extract of aerial parts of Euscaphis japonica (Tunb.) Kantiz (Staphyleaceae) showed significant inhibitory activity on HSC proliferation. Activity-guided fractionation led to the isolation of four triterpenoids, friedeline (1), glut-5-en-ol (2), pomolic acid (3), and methylrotundate (4). Among the triterpenoids isolated, pomolic acid (3) significantly inhibited the proliferation of HSCs at concentrations 10 and 100 μM.     

Genetic diversity and phylogenetic relationships in alder,Alnus firma, revealed by AFLP

Molecular markers for alder,Alnus firma Sieb. et Zucc, have not been studied extensively. Here, we used amplified fragment length polymorphism (AFLP) to investigate genetic relationships among 15 natural populations. EcoRI-ACG + Msel-CTG combinations revealed the highest polymorphism (62.2%). A total of 171 DNA fragments were identified. On average, 58.1% of the AFLP markers that were generated using four primer pairs were polymorphic. Diversity was insignificant among the populations. The combination of a wind-pollinated, outcrossing breeding system along with large population sizes, and the ability to regenerate by stump sprouting may explain the high level of genetic diversity within this species. The majority (98%) of the genetic variance resided within populations. The average number of individuals that were exchanged between populations per generation was very high (N _em = 12.3). Gene dispersal in alder is apparently by seed dispersalvia water and human activity as well as through pollen. Five individuals per population were claded in the same cluster.     

Two triterpenoids and other constituents from Petasites tricholobus

Two migrated ursane skeleton triterpenoids, one is D:B-friedoursane-3alpha,16alpha-dihydroxy-7alpha,8alpha-epoxy-5(10)-ene, named petatrichol A (1) and the other one represents a novel triterpenoid carbon framework, named petatrichol B (2), along with 10 known compounds were isolated from the rhizome of Petasites tricholobus. Their structures were elucidated on the basis of spectroscopic methods (IR, EIMS, HRMS, 1D and 2D NMR). The triterpenoids were assayed against Escherichia coli, Staphylococcus aureus and Bacillus subtilis. Compounds 1 and 2 exhibited significant antibacterial activity against B. subtilis.     

A novel synthetic oleanolic acid derivative (CPU-II<Subscript>2</Subscript>) attenuates liver fibrosis in mice through regulating the function of hepatic stellate cells

Regulation on the function of the hepatic stellate cells (HSCs) is one of the proposed therapeutic approaches to liver fibrosis. In the present study, we examined the in vitro and in vivo effects of CPU-II₂, a novel synthetic oleanolic acid (OLA) derivative with nitrate, on hepatic fibrosis. This compound alleviated CCl₄-induced hepatic fibrosis in mice with a decrease in hepatic hydroxyproline (Hyp) content and histological changes. CPU-II₂ also attenuated the mRNA expression of α-smooth muscle actin (α-SMA) and tissue inhibitor of metalloproteinase type 1 (TIMP-1) induced by CCl₄ in mice and reduced both mRNA and protein levels of α-SMA in HSC-T6 cells. Interestingly, CPU-II₂ did not affect the survival of HSC-T6 cells but decreased the expression of procollagen-α1 (I) in HSC-T6 cells through down-regulating the phosphorylation of p38 MAPK. Conclusion: CPU-II₂ attenuates the development of liver fibrosis rather by regulating the function of HSCs through p38 MAPK pathway than by damaging the stellate cells.     

Osthole ameliorates hepatic fibrosis and inhibits hepatic stellate cell activation

Background

Hepatic fibrosis is a dynamic process which ultimately leads to cirrhosis in almost patients with chronic hepatic injury. However, progressive fibrosis is a reversible scarring response. Activation of hepatic stellate cells (HSCs) is the prevailing process during hepatic fibrosis. Osthole is an active component majorly contained in the fruit of Cnidium monnieri (L.) Cusson. This present study investigated the therapeutic effects of osthole on rat liver fibrosis and HSC activation.

Results

We established the thioacetamide (TAA)-model of Sprague–Dawley (SD) rats to induce hepatic fibrosis. Rats were divided into three groups: control, TAA, and TAA + osthole (10 mg/kg). In vivo, osthole significantly reduced liver injury by diminishing levels of plasma AST and ALT, improving histological architecture, decreasing collagen and α-SMA accumulation, and improving hepatic fibrosis scores. Additionally, osthole reduced the expression of fibrosis-related genes significantly. Osthole also suppressed the production of fibrosis-related cytokines and chemokines. Moreover, nuclear translocation of p65 was significantly suppressed in osthole-treated liver. Osthole also ameliorated TAA-induced injury through reducing cellular oxidation. Osthole showed inhibitory effects in inflammation-related genes and chemokines production as well. In vitro, we assessed osthole effects in activated HSCs (HSC-T6 and LX-2). Osthole attenuated TGF-β1-induced migration and invasion in HSCs. Furthermore, osthole decreased TNF-α-triggered NF-κB activities significantly. Besides, osthole alleviated TGF-β1- or ET-1-induced HSCs contractility.

Conclusions

Our study demonstrated that osthole improved TAA-caused liver injury, fibrogenesis and inflammation in rats. In addition, osthole suppressed HSCs activation in vitro significantly.

Electronic supplementary material

The online version of this article (doi:10.1186/s12929-015-0168-5) contains supplementary material, which is available to authorized users.     

Pregnane, coumarin and lupane derivatives and cytotoxic constituents from Helicteres angustifolia

2alpha,7beta,20alpha-Trihydroxy-3beta,21-dimethoxy-5-pregnene (1), 6,7,9alpha-trihydroxy-3,8,11alpha-trimethylcyclohexo-[d,e]-coumarin (2), 3beta-hydroxy-27-benzoyloxylup-20(29)-en-28-oic acid (3), and 3beta-hydroxy-27-benzoyloxylup-20(29)-en-28-oic acid methyl ester (4), along with 24 known compounds were isolated and structurally characterized from roots and aerial parts of Helicteres angustifolia (Sterculiaceae). In a preliminary bioassay, the two cucurbitacin derivatives, cucurbitacin D and J exhibited significant inhibitory activities against the growth of both hepatocellular carcinoma BEL-7402 cells and malignant melanoma SK-MEL-28 cells in vitro.     

Peroxynitrite-mediated matrix metalloproteinase-2 activation in human hepatic stellate cells

To investigate whether hepatic stellate cells (HSCs) alter their expression of MMPs after exposure to nitrogen oxide intermediate (NOI), a human hepatic stellate cell line, LI90 cells, was stimulated with an NO donor, SNAP, or a peroxynitrite donor, SIN-1, and the culture supernatants were analyzed by gelatin zymography or anti-MMPs immunoblot. Although SIN-1 did not enhance the secretions of MMP-1 and 13, SIN-1 induced the NF-kappaB activation, MT1-MMP expression and the secretion of activated MMP-2 in LI90 cells. These results suggest that peroxynitrite may contribute to the remodeling of the extracellular matrix in liver by activating pro-MMP-2.     

Cytotoxic triterpenes from the aerial roots of Ficus microcarpa

Six triterpenes, 3beta-acetoxy-12,19-dioxo-13(18)-oleanene (1), 3beta-acetoxy-19(29)-taraxasten-20alpha-ol (2), 3beta-acetoxy-21alpha,22alpha-epoxytaraxastan-20alpha-ol (3), 3,22-dioxo-20-taraxastene (4), 3beta-acetoxy-11alpha,12alpha-epoxy-16-oxo-14-taraxerene (5), 3beta-acetoxy-25-methoxylanosta-8,23-diene (6) along with nine known triterpenes, 3beta-acetoxy-11alpha,12alpha-epoxy-14-taraxerene (7), 3beta-acetoxy-25-hydroxylanosta-8,23-diene (8), oleanonic acid (9), acetylbetulinic acid (10), betulonic acid (11), acetylursolic acid (12), ursonic acid (13), ursolic acid (14), and 3-oxofriedelan-28-oic acid (15) were isolated from the aerial roots of Ficus microcarpa, and their structures elucidated by spectroscopic methods. The in vitro cytotoxic efficacy of these triterpenes was investigated using three human cancer cell lines, namely, HONE-1 nasopharyngeal carcinoma, KB oral epidermoid carcinoma, and HT29 colorectal carcinoma cells. Compound 8 and pentacyclic triterpenes 9-15 possessing a carboxylic acid functionality at C-28 showed significant cytotoxic activities against the aforementioned cell lines and gave IC50 values in the range 4.0-9.4 microM.     

Antifungal constituents of the stem bark of Bridelia retusa

Antifungal activity guided fractionation of solvent extracts of the stem bark of Bridelia retusa of the family Euphorbiaceae against Cladosporium cladosporioides, furnished new bisabolane sesquiterpenes, (E)-4-(1,5-dimethyl-3-oxo-1-hexenyl)benzoic acid, (E)-4-(1,5-dimethyl-3-oxo-1,4-hexadienyl) benzoic acid, (R)-4-(1,5-dimethyl-3-oxo-4-hexenyl)benzoic acid and (-)-isochaminic acid, together with the known (R)-4-(1,5-dimethyl-3-oxohexyl)benzoic acid (ar-todomatuic acid), 5-allyl-1,2,3-trimethoxybenzene (elemicin), (+)-sesamin and 4-isopropylbenzoic acid (cumic acid). All these compounds showed fungicidal activity on TLC bioautography method at very low concentrations except elemicin.     

Diarylheptanoids from the rhizomes of Zingiber officinale

Seven new diarylheptanoids, i.e., (3S,5S)-3,5-diacetoxy-1,7-bis(4-hydroxy-3-methoxyphenyl)heptane, (3R,5S)-3-acetoxy-5-hydroxy-1,7-bis(4-hydroxy-3-methoxyphenyl)heptane, (3R,5S)-3,5-dihydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)heptane, (5S)-5-acetoxy-1,7-bis(4-hydroxy-3-methoxyphenyl)heptan-3-one, 5-hydroxy-1-(3,4-dihydroxy-5-methoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)heptan-3-one, 5-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-7-(3,4-dihydroxy-5-methoxy-phenyl)heptan-3-one and 1,5-epoxy-3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)heptane were isolated from the rhizomes of Chinese ginger (Zingiber officinale Roscoe), along with 25 known compounds, i.e., 8 diarylheptanoids, 14 gingerol analogs, a diterpene and 2 steroids. Their structures were elucidated by spectroscopic and chemical methods.     

Secondary metabolites from Commiphora opobalsamum and their antiproliferative effect on human prostate cancer cells

A cycloartane-type triterpenoid (1), an aliphatic alcohol glycoside (2), an eudesmane-type sesquiterpenoid (3), and a guaiane-type sesquiterpenoid (4) were isolated from the resinous exudates of Commiphora opobalsamum along with six known sesquiterpenoids (5-10). Their structures were established by extensive analysis of their 1D and 2D NMR spectroscopic data and chemical methods. The isolated compounds 1-3 and 5-9 were tested against human prostate cancer cell PC 3 and LNCaP. Among them, 1 and 2 showed moderate antiproliferative effects on human prostate cancer cell lines with IC50 values ranging from 5.7 to 23.6 microM; they were also able to inhibit the expression of androgen receptor (AR) in LNCaP cells. The six sesquiterpenoids were inactive in the bioassays.     

Antimicrobial constituents of Scrophularia deserti

A study of the chemistry and antibacterial activity of Scrophularia deserti led to the isolation of eight compounds, including the metabolite 3(zeta)-hydroxy-octadeca-4(E),6(Z)-dienoic acid (1). The known compounds ajugoside (2), scropolioside B (3), 6-O-alpha-L-rhamnopyranosylcatalpol (4), buddlejoside A(8) (5), scrospioside A (6), laterioside (7) and 3R-1-octan-3-yl-3-O-beta-D-glucopyranoside (8) were also isolated. Compounds 1-3 exhibited moderate antibacterial activity against strains of multidrug and methicillin-resistant Staphylococcus aureus (MRSA) and a panel of rapidly growing mycobacteria with minimum inhibitory concentration (MIC) values ranging from 32 to 128 microg/ml.     

Oxygenated diterpenes and other constituents from Moroccan Juniperus phoenicea and Juniperus thurifera var. africana

Six new diterpenic acids isolated as their methyl ester derivatives, i.e., methyl 12-oxo-8alpha,15-dihydroxyabiet-13-en-19-oate, methyl 12-oxo-8alpha-hydroxyabiet-13-en-19-oate, methyl 15-hydroperoxy-8alpha,12alpha-epidioxiabiet-13-en-19-oate, methyl 15-hydroxy-8alpha,12alpha-epidioxiabiet-13-en-19-oate, methyl 15-hydroperoxy-8alpha,14alpha,12alpha,13alpha-diepoxiabietan-13-en-19-oate, and methyl 7alpha,12beta-dihydroxysandaracopimarate, together with two new isovalerate derivatives of p-methoxycinnamyl alcohol and linalool, were isolated from the leaves of Juniperus thurifera var. africana and Juniperus phoenicea, grown in Morocco. The structures of these compounds were established by using spectroscopic techniques, including 2D NMR spectra. The cytotoxicity of the abietane diterpenoids was tested against five cell lines.     

Bioactive constituents of Artemisia monosperma

During a study on the chemistry and biological activity of Kuwaiti plants, new metabolites including 4,6-dihydroxy-3-[3'-methyl-2'-butenyl]-5-[4'-hydroxy-3'-methyl-2'-butenyl]-cinnamic acid (1), the 3R,8R stereoisomer of the C17 polyacetylene dehydrofalcarindiol (2) and a C10 polyacetylene glucoside (3) were characterised by spectroscopic means. Additionally, the previously characterised natural products 1,3R,8R-trihydroxydec-9-en-4,6-yne (4), spathulenol (5) and eriodyctiol-7-methyl ether (6) were also isolated. Compounds 2, 3, and 4 were evaluated for their ability to inhibit the enzyme 12-lipoxygenase and 3 and 4 showed moderate activity at 30 microg/ml. Compound 2 was evaluated against a panel of colorectal and breast cancer cell lines and IC50 values ranged from 5.8 to 37.6 microg/ml. Against a panel of fast-growing mycobacteria and a standard ATCC strain of Staphylococcus aureus, compound 6 exhibited minimum inhibitory concentrations in the range of 64-128 microg/ml.     

Bioactivity guided isolation of anticancer constituents from leaves of Alnus sieboldiana (Betulaceae)

The leaves of the Japanese Alnus sieboldiana have been extracted with n-hexane and then with methanol. A bioactivity-guided approach based on MTT assay for growth inhibition and quantitative real-time PCR for TNF-α inhibitory activity was taken to identify the active compounds in EtOAc soluble fraction of the methanol extract. From this active fraction, seven compounds have been isolated and four compounds (pinosylvin, galangin, quercetin and methyl gallate) have been examined for their dose-response effect on the viability of A549 cells and on TNF-α inhibitory activity. Based on MTT assay, all of the four examined compounds inhibit growth of human lung cancer cells. Among four tested compounds only galangin (3,5,7-trihydroxyflavone) significantly inhibited TNF-α gene expression in A549 cells (IC₅₀ = 94 μM). Taken together, this finding suggests that galangin may be useful in cancer prevention.     

Immunosuppressive constituents from Saussurea medusa.

The methanol extract of Saussurea medusa Maxim afforded two lignans: 2alpha-guaicyl-4-oxo-6alpha-catechyl-3,7-dioxabicyclo [3.3.0]octane and 1alpha-hydroxy-2alpha,4alpha-guaicyl-3,7-dioxabicyclo[3.3.0]octane; two chlorophyll derivatives: 13-epi-phaeophorbide-a and 13-epi-phaeophorbide-a methyl ester; one megastigmane derivative: 3beta-hydroxy-5alpha,6alpha-epoxy-7-megastigmen-9-one, along with 19 known compounds. Their structures were established on the basis of spectroscopic studies.     

Chemical constituents of Picea neoveitchii

Four flavonoids, 5,7,4′-trihydroxy-3,8,-dimethoxy-6-C-methylflavone (1), 5,8,4′-trihydroxy-3,7-dimethoxy-6-C-methylflavone (2), 7-methoxy-6-C-methylkaempferol (3) and kaempferol-7-O-(2″-E-p-coumaroyl)-α-l-arabinofuranoside (4), together with 15 known compounds, were isolated from the twigs and leaves of Picea neoveitchii Mast. Their structures were elucidated on the basis of analyses of spectroscopic data. Compound 4 showed strong anti-fungal activity against Fusarium oxysporum whereas compounds 1-4 were all active against Rhizoctonia solani.     

Eudesmane derivatives and other constituents from Saussurea parviflora

A survey of the whole plant of Saussurea parviflora afforded three compounds 11,12,13-trihydroxy-4(15),8-eudesmadiene-9-one, eudesman-8beta,12-olide-1-O-beta-D-glucoside and 1beta,3beta-dihydroxyursa-9(11),12-diene-3-octadecanoate, as well as 13 known compounds. Their structures were elucidated on the basis of spectral evidence, especially by using NMR spectroscopic techniques. In addition, encelin exhibited effective antitumor activity on L02, SMMC-7721 and HO-8910 cells.