Objective:To investigate the molecular mechanism underlying the anti-hepatic fibrosis activity of ethyl acetate fraction Dicliptera chinensis(L.)Juss.(EDC)in human hepatic stellate cells(HSCs)in vitro and in a carbon ...Objective:To investigate the molecular mechanism underlying the anti-hepatic fibrosis activity of ethyl acetate fraction Dicliptera chinensis(L.)Juss.(EDC)in human hepatic stellate cells(HSCs)in vitro and in a carbon tetrachloride(CCl4)-induced hepatic fibrosis mouse model in vivo.Methods:For in vitro study,HSCs were pre-treated with platelet-derived growth factor(10 ng/mL)for 2 h to ensure activation and treated with EDC for 24 h and 48 h,respectively.The effect of EDC on HSCs was assessed using cell counting kit-8 assay,EdU staining,transmission electron microscopy,immunofluorescence staining,and Western blot,respectively.For in vivo experiments,mice were intraperitoneally injected with CCl4(2μL/g,adjusted to a 25%concentration in olive oil),3 times per week for 6 weeks,to develop a hepatic fibrosis model.Forty 8-week-old male C57 BL/6 mice were divided into 4 groups using a random number table(n=10),including control,model,positive control and EDC treatment groups.Mice in the EDC and colchicine groups were intragastrically administered EDC(0.5 g/kg)or colchicine(0.2 mg/kg)once per day for 6 weeks.Mice in the control and model groups received an equal volume of saline.Biochemical assays and histological examinations were used to assess liver damage.Protein expression levels ofα-smooth muscle actin(α-SMA)and microtubule-associated protein light chain 3 B(LC3 B)were measured by Western blot.Results:EDC reduced pathological damage associated with liver fibrosis,downregulated the expression ofα-SMA and upregulated the expression of LC3 B(P<0.05),both in HSCs and the CCl4-induced liver fibrosis mouse model.The intervention of bafilomycin A1 and rapamycin in HSCs strongly supported the notion that inhibition of autophagy enhancedα-SMA protein expression levels(P<0.01).The results also found that the levels of phosphoinositide(PI3 K),p-PI3 K,AKT,p-AKT,mammalian target of rapamycin(mTOR),p-mTOR,and p-p70 S6 K all decreased after EDC treatment(P<0.05).Conclusion:EDC has anti-hepatic fibrosis activity by inducing au展开更多
[Objectives] To study the acute toxicity,mutation,and feeding experiment of polysaccharides from Dicliptera chinensis( L.)Juss.,and assess its safety. [Methods]In accordance with Procedures for toxicological Assessmen...[Objectives] To study the acute toxicity,mutation,and feeding experiment of polysaccharides from Dicliptera chinensis( L.)Juss.,and assess its safety. [Methods]In accordance with Procedures for toxicological Assessment on Food Safety,acute toxicity to mice,micronucleus experiment for bone marrow cell in mice,sperm shape abnormality test in mice,and Ames experiment were carried out,and 30 d feeding experiment was detected for rats. [Results] when the maximum tolerance dose( MTD) of mice was higher than 20 g/kg,at polysaccharides from Dicliptera chinensis( L.) Juss. ≤10 g/kg,there was no statistical difference between the control group and mouse bone marrow cell micronucleus and sperm shape abnormality test group; Ames experiment indicated that the number of revertant colonies of different dose groups of polysaccharides from Dicliptera chinensis( L.) Juss. with and without S9 did not exceed two times of the blank control group; rat 30 d feeding experiment indicated that when polysaccharides from Dicliptera chinensis( L.) Juss. ≤ 3. 2 g/kg,rats grew and developed well,and there was no statistical difference with the control group in blood,multiple organ biochemical indicators and liver tissue. [Conclusions]Polysaccharides from Dicliptera chinensis( L.) Juss. are safe and non-toxic chemical compounds and can be used as health products or drugs.展开更多
基金Supported by the National Natural Science Foundation of China(No.81673774)the Administration of Traditional Chinese Medicine of Shandong Province(2019-0447)。
文摘Objective:To investigate the molecular mechanism underlying the anti-hepatic fibrosis activity of ethyl acetate fraction Dicliptera chinensis(L.)Juss.(EDC)in human hepatic stellate cells(HSCs)in vitro and in a carbon tetrachloride(CCl4)-induced hepatic fibrosis mouse model in vivo.Methods:For in vitro study,HSCs were pre-treated with platelet-derived growth factor(10 ng/mL)for 2 h to ensure activation and treated with EDC for 24 h and 48 h,respectively.The effect of EDC on HSCs was assessed using cell counting kit-8 assay,EdU staining,transmission electron microscopy,immunofluorescence staining,and Western blot,respectively.For in vivo experiments,mice were intraperitoneally injected with CCl4(2μL/g,adjusted to a 25%concentration in olive oil),3 times per week for 6 weeks,to develop a hepatic fibrosis model.Forty 8-week-old male C57 BL/6 mice were divided into 4 groups using a random number table(n=10),including control,model,positive control and EDC treatment groups.Mice in the EDC and colchicine groups were intragastrically administered EDC(0.5 g/kg)or colchicine(0.2 mg/kg)once per day for 6 weeks.Mice in the control and model groups received an equal volume of saline.Biochemical assays and histological examinations were used to assess liver damage.Protein expression levels ofα-smooth muscle actin(α-SMA)and microtubule-associated protein light chain 3 B(LC3 B)were measured by Western blot.Results:EDC reduced pathological damage associated with liver fibrosis,downregulated the expression ofα-SMA and upregulated the expression of LC3 B(P<0.05),both in HSCs and the CCl4-induced liver fibrosis mouse model.The intervention of bafilomycin A1 and rapamycin in HSCs strongly supported the notion that inhibition of autophagy enhancedα-SMA protein expression levels(P<0.01).The results also found that the levels of phosphoinositide(PI3 K),p-PI3 K,AKT,p-AKT,mammalian target of rapamycin(mTOR),p-mTOR,and p-p70 S6 K all decreased after EDC treatment(P<0.05).Conclusion:EDC has anti-hepatic fibrosis activity by inducing au
基金Supported by Project of National Natural Science Foundation(81360685)Traditional Chinese Medicine Science and Technology Planning Project of Health and Family Planning Commission of Guangxi Zhuang Autonomous Region(GZBZ16-15)
文摘[Objectives] To study the acute toxicity,mutation,and feeding experiment of polysaccharides from Dicliptera chinensis( L.)Juss.,and assess its safety. [Methods]In accordance with Procedures for toxicological Assessment on Food Safety,acute toxicity to mice,micronucleus experiment for bone marrow cell in mice,sperm shape abnormality test in mice,and Ames experiment were carried out,and 30 d feeding experiment was detected for rats. [Results] when the maximum tolerance dose( MTD) of mice was higher than 20 g/kg,at polysaccharides from Dicliptera chinensis( L.) Juss. ≤10 g/kg,there was no statistical difference between the control group and mouse bone marrow cell micronucleus and sperm shape abnormality test group; Ames experiment indicated that the number of revertant colonies of different dose groups of polysaccharides from Dicliptera chinensis( L.) Juss. with and without S9 did not exceed two times of the blank control group; rat 30 d feeding experiment indicated that when polysaccharides from Dicliptera chinensis( L.) Juss. ≤ 3. 2 g/kg,rats grew and developed well,and there was no statistical difference with the control group in blood,multiple organ biochemical indicators and liver tissue. [Conclusions]Polysaccharides from Dicliptera chinensis( L.) Juss. are safe and non-toxic chemical compounds and can be used as health products or drugs.
