Effects of ethanolic extracts of Euonymus alatus on CCl / 中国药理学通报

Aim To explore the mechanism of ethanolic extracts of euonymus alatus on CCl4-induced hepatic fibrosis in mice by regulating JAK2/STAT3 signaling pathway. Methods Sixty C57BL/6J mice were randomly divided into control group,model group,EAL,EAM),EAH,and Silybin(n=10). Except for the control group,mice in other groups were injected with 25% CCl4 of 1.6 mL·kg

Prevention of liver fibrosis in mice by ethanol extract of Euonymus alatus and its preliminary mechanism / 中华消化杂志

Objective:To study the role of ethanol extract of Euonymus alatus stems (EAT) and ethanol extract of Euonymus alatus wings (EAW) in anti-hepatic fibrosis induced by carbon tetrachloride in mice, and to explore its preliminary mechanism. Methods:Sixty C57BL/6 mice were selected and randomly divided into healthy control group, carbon tetrachloride model (CTM) group, EAW low dose (EAW-L) group, EAW high dose (EAW-H) group, EAT low dose (EAT-L) group and EAT high dose (EAT-H) group, with 10 mice in each group. Three days before modeling, the mice of EAT-L, EAT-H, EAW-L and EAW-H group were gavaged with EAT or EAW at 2.0 or 8.0 g/kg, respectively, and the mice of healthy control group and CTM group were gavaged with equal volume of pure water, once a day till the 30th day after modeling (total 33 times). Five percent carbon tetrachloride olive oil solution was intraperitoneally injected at 8 mL/kg to establish liver fibrosis model in CTM, EAT-L, EAT-H, EAW-L and EAW-H groups. The mice in the healthy control group were intraperitoneally injected with equal volume of 0.9% sodium chloride solution, twice per week for 30 days, and a total of 9 times of injection. The liver index, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil) and interleukin-6 (IL-6) were detected. Hematoxylin-eosin and Masson staining were used to observe the pathological changes of mouse liver tissue and calculate the collagen volume fraction. The liver inflammatory response and fibrosis degree were evaluated by histological activity index (HAI) and Ishak system score. The level of α-smooth muscle actin(α-SMA)in liver tissue was both detected by immunohistochemistry and Western blotting. The expression of matrix metalloproteinase 2 (MMP2) and extracellular signal-regulated kinase (ERK) 1/2 at protein and mRNA level was detected by Western blotting and fluorescent quantitative polymerase chain reaction. Analysis of variance, Tukey test and Dunn test were used for statistical analysis.Results:The hepatic indexes of EAW-L, EAW-H and EAT-H groups were lower than that of CTM group(0.06±0.01, 0.05±0.01 and 0.05±0.01 vs. 0.07±0.01), and the differences were statistically significant ( q=5.12, 7.70, 7.11; all P<0.01). The serum ALT and AST levels of EAW-L, EAW-H, EAT-L and EAT-H groups were lower than those of CTM group((601.76±141.38), (283.35±42.32), (734.74±116.06) and (391.60±34.33) U/L vs.(982.45±96.04) U/L, (509.49±152.29), (345.41±67.39), (282.30±65.72) and(243.23±45.20) U/L vs.(766.01±114.49) U/L), and the differences were statistically significant ( qALT =9.88, 20.81, 7.65, 17.58, qAST =5.11, 12.52, 14.92, 15.56; all P<0.001). The serum TBil levels of EAW-H and EAT-H groups were lower than that of CTM group((6.81±0.49) and (7.08±1.78) μmol/L vs.(12.68±3.28) μmol/L), and the differences were statistically significant( q=6.31, 6.01; both P<0.01). The serum IL-6 levels of EAW-L, EAW-H, EAT-L and EAT-H groups were lower than that of CTM group((29.26±5.42), (24.28±4.75), (9.05±1.74) and (8.01±1.24) ng/L vs.(53.21±10.05) ng/L); the serum IL-6 level of EAT-L group was lower than that of EAW-L group; the serum IL-6 level of EAT-H group was lower than that of EAW-H group, and the differences were statistically significant( q=12.20, 14.73, 22.48, 22.11, 10.28, 7.96; all P <0.001). The collagen volume fractions of EAW-L, EAW-H, EAT-L and EAT-H groups were lower than that of CTM group (6.15±1.09, 2.91±0.76, 7.07±1.37 and 5.31±0.80 vs. 12.36±1.96); the collagen volume fraction of EAW-H group was lower than that of EAW-L, EAT-L and EAT-H groups, and the differences were statistically significant( q=11.68, 17.78, 9.94, 13.25; 6.10, 7.84, 4.53; all P <0.05). The HAI and Ishak system scores of EAW-H and EAT-H groups were lower than those of CTM group (6.0 (5.5, 7.5) and 7.0 (6.0, 7.5) vs. 13.0 (12.0, 13.0), 1.0 (1.0, 2.0) and 2.0 (1.0, 2.0) vs. 4.0 (3.0, 4.0)), and the differences were statistically significant( ZHAI=3.38, 3.23, Zlshak=3.22, 3.03; all P<0.05). The result of immunohistochemical analysis showed that the expression levels of α-SMA in the mice liver tissues of EAW-L, EAW-H, EAT-L, EAT-H and CTM groups were 4.76±0.36, 2.75±0.29, 3.72±0.34, 5.20±0.79 and 5.98±0.52, respectively. The result of Western blotting showed that the expression levels of α-SMA in the mice liver tissues of CTM, EAW-L, EAW-H, EAT-L and EAT-H groups were 0.96±0.11, 0.67±0.07, 0.22±0.01, 0.78±0.08 and 0.68±0.07, respectively. Two detection methods both showed that the expression levels of α-SMA of EAW-L, EAW-H and EAT-H groups were lower than that of CTM group; the expression level of α-SMA of EAW-H group was lower than that of EAW-L, EAT-L and EAT-H group, and the differences were statistically significant( qimmunohistochemical =6.06, 15.95, 11.18, 9.92, 12.10 and 4.79, qWestern blotting=7.29, 18.34, 6.84, 11.05, 13.97 and 11.49, all P<0.05). The expression levels of MMP2 and ERK1/2 at protein and mRNA levels in the mice liver tissues of EAW-L, EAW-H, EAT-L, EAT-H and CTM groups were 0.18±0.04, 0.16±0.04, 0.28±0.02, 0.21±0.02 and 0.84±0.02, 0.80±0.02, 0.57±0.08, 0.83±0.03, 0.69±0.02 and 0.91±0.04, 18.74±1.90, 10.73±1.24, 24.99±1.84, 7.19±0.48 and 24.68±1.18, 29.44±4.47, 11.96±0.53, 24.75±4.04, 5.30±0.36 and 35.76±0.85, respectively. The expression levels of MMP2 at protein level in EAW-L, EAW-H, EAT-L and EAT-H groups were lower than that in CTM group; the expression levels of ERK1/2 at protein level in EAW-H and EAT-H groups were lower than that in CTM group; the expression level of ERK1/2 at protein level in EAW-H group was lower than that in EAT-H group; the expression levels of MMP2 and ERK1/2 at mRNA level in EAW-H and EAT-H group were lower than those in CTM group; the expression levels of MMP2 and ERK1/2 at mRNA level in EAW-H group were lower than those in EAW-L group; the expression levels of MMP2 and ERK1/2 at mRNA level in EAT-H group were lower than those in EAT-L and EAW-H groups, and the differences were statistically significant( q=22.15, 22.96, 18.87, 21.31; 13.42, 8.53; 4.90; 18.57, 23.29, 16.49, 21.11; 10.66, 12.12; 23.70, 15.38, 13.48, 16.73; all P<0.05). Conclusions:Both EAT and EAW can alleviate carbon tetrachloride-induced liver injury and liver fibrosis in mice, which may be related with inhibiting the expression of ERK1/2 and IL-6 and then affecting the Ras/ERK-MMP2 signaling pathway.

Chemistry research on natural product derivatives of dihydro-β-agarofuran sesquiterpenoids / 中国中药杂志

Dihydro-β-agarofuran sesquiterpenoids possess chemical diversity and biodiversity. A dihydro-β-agarofuran sesquiterpenoid with only hydroxyl groups has been prepared by basic hydrolysis of crude extract of Euonymus bungeanus with 0.4% yield. Twelve derivatives were available in esterification, oxidation, decarboxylation, etc. Extensive ~1H-NMR,~(13)C-NMR, MS spectroscopic analyses and single-crystal X-ray diffraction analysis with Cu Kα radiation indicated that eleven derivatives were new compounds. The results will provide reference for chemistry study on natural product derivatives of dihydro-β-agarofuran sesquiterpenoids.

The inhibitory effect of Euonymus alatus against mouse hepatic fibrosis induced by CCl4 / 中国药理学通报

Aim To explore the inhibitory effect of Euonymus alatus on hepatic fibrosis induced by carbon tetrachloride (CCl4) in mice and its mechanism. Methods Eighty C57BL/6 male mice were randomly divided into eight groups: normal group, CCl4model group, Euonymus alatus(EA) ethanol extracts groups in early stage(EAE), EA ethanol extracts groups in later stage(EAL),two drug groups with low/medium/high dose(EAE-L/M/H, EAL-L/M/H), with 10 mice in each group. Fibrosis model was established by injecting CCl4in peritoneal cavity,and the study lasted for 30 days. Different doses of drugs were given from 1 st day to 15 th day in EAE while from 16 th day to 30 th day in EAL,then all mice were sacrificed to for the observation of the morphological changes and collage-nous fiber by HE and Masson staining. Liver index, ALT,AST and TNF-α were tested by ELISA. The ex-pressions of α-SMA and CollagenⅠwere measured by immunohistochemistry and Western blot. Results Compared to normal group, liver index, ALT, AST, TNF-α, α-SMA and CollagenⅠ in EA groups were lower than those in model group in a dose-dependent manner(P<0.01 or P<0.05). Liver morphology and collagenous fiber in EAE and EAL were better than those in model group in a dose-dependent manner. The effect of EAE were superior to that of the EAL in HE, Masson, α-SMA, Collagen Ⅰ indexes(P <0.05). Conclusions Euonymus alatus may inhibit the process of hepatic fibrosis in mice with dose-effect de-pendence, and drug treatment in early stage performs better,which may be related to the decrease of TNF-α that affects the expression of α-SMA and Collagen Ⅰ.

Study on Chemical Constituents of Ethanol Extract from Euonymus amygdalifolius / 中国药房

OBJECTIVE:To study the chemical constituents of Euonymus amygdalifolius.METHODS:Silica gel column chromatogram,gel column chromatogram,TLC and semi-preparative HPLC were adopted to isolate and purify ethanol extract from E.amygdalifolius.The structure of compounds was analyzed and identified according to physical and chemical properties,spectral data (mass spectrurn,hydrogen spectrum and carbon spectrum).RESULTS:Ten compounds were isolated from ethanol extract of aerial part of E.amygdalifolius,i.e.taraxerol (1),sophoradiol (2),taraxerone (3),sorghumol (4),heptaecanoic acid (5),octadecenoic acid (6),β-Sitosterol (7),daucosterol (8),epifriedelinol (9) and friedelin (10).CONCLUSIONS:Above compounds are all isolated from E.amygdalifolius for the first time;compounds 1,2,4,5,6 are isolated and obtained from Euonymus A.for the first time.The study lay a foundation for quality evaluation of E.amygdalifolius.

A new cembranoid diterpene from twigs of Euonymus alatus / 中草药

Objective To study the chemical constituents from the twigs of Euonymus alatus. Methods Compounds were isolated and purified by a combination of various chromatographic techniques, and their structures were elucidated by physiochemical property and spectral analysis. Results Three compounds were obtained from 95% ethanol extract of the twigs of E. alatus and identified as (1S,2S,7E,11R,12S)-2,11-dihydroxy-1,12-oxidocembra-4(18),7(8)-diene (1), hemerocallal A (2), and betulinic acid methyl ester (3). Conclusion Compound 1 is a new cembranoid diterpene and named euonymuditerpene A, and compounds 2 and 3 are isolated from the twigs of Euonymus alatus for the first time.

Protective effect and mechanism of serum containing Euonymus fortunei on rat pancreatic islet cells / 器官移植

Objective To investigate the protective effect and mechanism of serum containing Euonymus fortunei on the rat pancreatic islet cells. Methods Forty male SD rats were randomly divided into 5 groups (n=8 in each group), including the control group (normal rat islet cells were cultured with normal rat serum), ischemic preconditioning group (abdominal aorta was blocked first and then re-opened before the pancreas was obtained, and the pancreatic islet cells were cultured with normal rat serum), Euonymus fortunei treatment group (normal rat islet cells were cultured with rat serum containing Euonymus fortunei), Euonymus fortunei group and blank group (normal rats were administered orally with Euonymus fortunei extract or distilled water for the preparation of rat serum). Diphenylthiocarbazone (DTZ) staining was utilized to observe and calculate the quantity of islets. Acridine orange (AO)/propidium iodide (PI) staining was adopted to calculate the survival rate of islet cells. The insulin release experiment was performed to calculate the stimulation index (SI) and evaluate islet cell function. The concentration of glutathione (GSH) and nitric oxide (NO) in islet cells was detected using GSH and NO kits. The expression level of inducible nitric oxide synthase (iNOS) messenger RNA (mRNA) was quantitatively measured by reverse transcription polymerase chain reaction (RT-PCR). Results Islet cells were observed in specifically scarlet color after DTZ staining. The quantity of islet cells did not significantly differ among different groups (all P>0.05). Along with the prolongation of culture time, the activity of islet cells in each group was gradually decreased. At 72 h after isolation and culture, compared with the control group, the survival rate of the cells was significantly higher in the Euonymus fortunei treatment group (P<0.05). The insulin release test results demonstrated that compared with the control group, the SI of the ischemic preconditioning and Euonymus fortunei treatment groups was significantly increased (both P<0.05). Compared with the control group, the GSH contents of pancreatic islet cells in the ischemic preconditioning and Euonymus fortunei treatment groups were considerably enhanced, the NO content was significantly decreased, and the expression level of iNOS mRNA was significantly down-regulated (all P<0.05). Conclusions Euonymus fortunei can increase the survival rate of islet cells and enhance the function of pancreatic islets by increasing the level of GSH, down-regulating the expression of iNOS and decreasing the NO production.

Protective Effect of the Extract of Euonymus Hederaceuson on H2O2-induced Oxidative Damage of Vascular Endothelial Cells / 中国药师

Objective: To observe the protective effects of the alcohol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceuson on H2O2-induced oxidative damage of vascular endothelial cells ECV-304.Methods: ECV-304 cells were cultured in vitro and H2O2-induced oxidative damage in ECV-304 cells was used as the model.After the treatment with ethanol extract, ethyl acetate extract or 1-butanol extract of Euonymus hederaceus (the final concentration of 400, 200, 100, 50 and 25 μg·ml-1) for 24h, the cell viability was detected by MTT assay.Hoechst staining was used to observe the cell apoptosis, and then the supernatant was collected, and the contents of SOD, NOS and NO were used to determine the effects of the extracts on the oxidative stress of vascular endothelial cells.Results: The ethanol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceus within the concentration range of 25-400 μg·ml-1 showed no cytotoxicity on ECV-304 cells, and the ethyl acetate extract at the concentration of 400μg·ml-1 had certain proliferation effect on the cells (P<0.05).The ethyl acetate extract at the concentration of 50-400 μg·ml-1 and the 1-butanol extract at the concentration of 100-400 μg·ml-1of Euonymus hederaceus could increase the survival rate of ECV-304 cells with oxidative injury significantly, and the OD value showed significant difference when compared with that of the model group (P<0.05 or P<0.01).The ethanol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceus could significantly improve the SOD and NO levels in ECV-304 cells with oxidative damage (P<0.05 or P<0.01), and had no notable influence on NOS level.The ethanol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceus could inhibit the cell apoptosis with oxidative damage and improve the nuclear morphology, and the apoptosis exhibited significant difference when compared with that of the model group (P<0.01).Conclusion: The extracts of Euonymus hederaceus have protective effects on vascular endothelial cells with oxidative damage induced by H2O2.

Dihydrobenzofuran Neolignans Isolated from Euonymus alatus Leaves and Twigs Attenuated Inflammatory Responses in the Activated RAW264.7 Macrophage Cells

Anti-inflammatory effects of dihydrobenzofuran neolignans isolated from Euonymus alatus leaves and twigs were evaluated in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Six neolignans, (+)- simulanol (1), (+)-dehydrodiconiferyl alcohol (2), (-)-simulanol (3), (-)-dehydrodiconiferyl alcohol (4), (+)-dihydrodehyrodiconiferyl alcohol (5), threo-buddlenol B (6) effectively inhibited the production of nitric oxide (NO) induced by LPS, and the activity of iNOS. (-)-dehydrodiconiferyl alcohol (4), which showed the most potent inhibitory activity, attenuated the activity of iNOS enzyme and also the expression of iNOS and COX-2 proteins. The subsequent production of pro-inflammatory cytokines, interleukin-1β, interleukin-6, tumor necrosis factor-α and prostaglandin E2 were also inhibited by the pretreatment of RAW264.7 cells with (-)-dehydrodiconiferyl alcohol (4). These neolignans are thought to contribute to anti-inflammatory effects of E. alatus, and expected to be potential candidates to prevent/treat inflammation-related diseases.

Inhibitory activity of Euonymus alatus against alpha-glucosidase in vitro and in vivo

The major goal in the treatment of diabetes mellitus is to achieve near-normal glycemic control. To optimize both fasting blood glucose and postprandial glucose levels is important in keeping blood glucose levels as close to normal as possible. alpha-Glucosidase is the enzyme that digests dietary carbohydrate, and inhibition of this enzyme could suppress postprandial hyperglycemia. The purpose of this study was to test the inhibitory activity of methanol extract of Euonymus alatus on alpha-glucosidase in vitro and in vivo to evaluate its possible use as an anti-diabetic agent. Yeast alpha-glucosidase inhibitory activities of methanol extract of E. alatus were measured at concentrations of 0.50, 0.25, 0.10, and 0.05 mg/ml. The ability of E. alatus to lower postprandial glucose was studied in streptozotocin-induced diabetic rats. A starch solution (1 g/kg) with and without E. alatus extract (500 mg/kg) was administered to diabetic rats by gastric intubation after an overnight fast. Plasma glucose levels were measured at 30, 60, 90, 120, 180, and 240 min. Plasma glucose levels were expressed in increments from baseline, and incremental areas under the response curve were calculated. Extract of E. alatus,which had an IC50 value of 0.272 mg/ml, inhibited yeast alpha-glucosidase activity in a concentration-dependent manner. A single oral dose of E. alatus extract significantly inhibited increases in blood glucose levels at 60 and 90 min (p<0.05) and significantly decreased incremental response areas under the glycemic response curve (p<0.05). These results suggest that E. alatus has an antihyperglycemic effect by inhibiting alpha-glucosidase activity in this animal model of diabetes mellitus.

Chemical constituents in active fraction from Euonymus alatus with anti-myocardial ischemic effect / 中草药

Objective To study the chemical constituents of the EtOAc soluble fraction obtained from the stems and branches of Euonymus alatus with anti-myocardial ischemic effect. Methods Chemical constituents were isolated and purified by solvent extraction together with various chromatographic techniques, and the structures were identified by physicochemical properties and spectral analyses. ResultsFive compounds, 1-[3-?-D-glucopyranosyloxy)-4, 5-dihydroxyphenyl]-ethanone (Ⅰ), phenethyl alcohol 8-O-?-D-glucopyranosyl-(1→2)-?-D-glucopyranoside (Ⅱ), eugenyl-O-?-D-glucopyranoside (Ⅲ), hyperin (Ⅳ), and hesperidin (Ⅴ) were isolated. Conclusion Compound Ⅰ is determined as a new compound named guijianyuside. Compounds Ⅱ and Ⅲ are isolated from this plant for the first time.

Chemical constituents from wing twigs of Euonymus alatus / 中草药

Objective To study the chemical constituents from the wing twigs of Euonymus alatus. Methods Compounds were isolated and purified repeatedly on silica gel, Sephadex LH-20, open RP-C18 column chromatographies and PTLC, and their chemical structures were elucidated by their physicochemical properties and spectral data, such as 1H-NMR, 13C-NMR, and EI-MS. Results Nine compounds were obtained and identified as: epifriedelinol (Ⅰ), stigmast-4-en-3-one (Ⅱ), 6?-hydroxy-stigmast-4-en-3-one (Ⅲ), ?-sitosterol (Ⅳ), methyl 2, 4-dihydroxy-3, 6-dimethyl benzoate (Ⅴ), methyl 2, 4-dihydroxy-6-methyl benzoate (Ⅵ), 7-methoxy-4-methylphthalide (Ⅶ), vanillin (Ⅷ), n-octacosanol (Ⅸ). Conclusion Compound Ⅶ is first reported as a natural product. Compounds Ⅴ-Ⅷ are reported from plants of Euonymus L. for the first time.

Quality Standard of Tangkang Capsule / 中国药房

OBJECTIVE:To establish the quality standard of Tangkang capsule. METHODS:Momordica charantia L. and Euonymus alatus(Thunb.)Sieb.,the main ingredients in Tangkang capsule including were identified by TLC, meanwhile, the content of total Momordica charantian saponin was determined by UV-Vis. RESULTS: The TLC spots were clear and the separation degree was good. The linear range of Ginsenoside Rg1 was 72～252 ?g(r=0.999 7) with average recovery rate at 100.68% (RSD=1.43%,n=6). CONCLUSION: The established quality standard can be used for the quality control of this Tangkang capsule.

An experimental research on inhibition of immediate and delayed type hypersensitivity by the 70% ethanolic extract from euonymus alatus / 中国药理学通报

ABSTRACT AIM To explore the restrained activities of the 70% ethanolic extract from euony-mus alatus(EAL) on the immediate and the early and late phases of delayed type hypersensitivity (DTH), and explore its mechanisms of action. METHODS EAL was pre-incubated with rat peritoneal mast cells and isolated cavia porcellus ile-um, respectively. Its effects on histamine release were elicited by compound 48/80 and ileum constriction by 5-PC. Obserseved the effects of EAL, which separated total flavanid from, it HT were observed. In mouse DTH models upon challenge with SRBC or was on the early and late phases by administration after antigen challenge. RESULTS EAL had a dependent restraint-effect to release the histamine from rat peritoneal mast cells and showed a restrained tendency on the cavia porcellus ileum constriction. Meantime, EAL and its total flavanid can restrain the two phases of DTH too. Which were stronger on the late phase than early. CONCLUSION The steroid and flavanid of EAL can restrain the early and late phase of DTH by stabling mast cell membrane and restraing histamine and 5-HT release. Its flavanid is the active component to restrain DTH.