The Effect Of Dendrobium Officinale Different Extracts On TGFβ1-induced HSC-T6 Cell Line

Objective:To investigate the effects of the Dendrobium officinale different extracts on the proliferation and expression of cytokines of TGFβ1-induced HSC-T6 cell line,and to explore the different anti-hepatic fibrosis functions of polysaccharide,methanol extracts and different extraction parts of methanol extraction.Methods:(1)MTT method was used to evaluate the effects of different extracts of Dendrobium officinale on HSC-T6 proliferation,with inhibition rate within 10%of the concentration is non-toxic concentration.TGFβ1 was used to induce HSC-T6 cells to establish the proliferation model,then cells were divided into blank control group,model group,polysaccharide group and methanol extract group.MTT assay was applied to observe the dose or time dependant effect of extracts on TGFβ1-induced HSC-T6 cells proliferation;ELISA assay was used to evaluate the influence of polysaccharide and methanol extract on the content of COL-Ⅲ,COL-Ⅳ,α-SMA,CTGF and PDGF in HSC-T6 cells supernatant;(2)The methanol extract of Dendrobium officinale was divided into n-hexane layer,chloroform layer,ethyl acetate layer,n-butanol layer and water layer which were used to evaluate forther.After measuring dose-effect of each layer extracts on TGFβ1-induced HSC-T6 cells;a fluorescence RT-PCR assay was emplyed to measure the effect of each layer of methanol extracts on expression of α-SMA,CTGF,Endoglin,TβR-Ⅱ and Caspase3 mRNA in HSC-T6 cells;ELISA assay was applied to detect the influence of each layer of methanol extracts on the content of α-SMA,CTGF,Endoglin,TβR-Ⅱ and Caspase3 in HSC-T6 cells ssupernatant.Results:(1)MTT assay results showed that both aqueous and methanol extract had inhibitory effects on the proliferation of TGFβ1-induced HSC-T6 cells,neverthless,there was no significant difference found on their inhibition performances.Despite ELISA aasay results showed both aqueous and methanol extract had some inhibitory effects on α-SMA,COL-Ⅲ,CTGF and PDGF,their inhibition performances differences were not significant;(2)MTT assay results showed that not only n-hexane group,chloroform group,ethyl acetate group had an inhibitory effects on the proliferation of TGFβ1-induced HSC-T6 cells,but also there were sinificant differences found among the groups(P<0.05).RT-PCR,ELISA aasay results showed that compared with the blank control group,model group increased the mRNA and protein content of α-SMA、CTGF(P<0.05),Caspase3,decreased Endoglin、TβR-Ⅱ(P>0.05).N-hexane group reduced the content of α-SMA,CTGF,TβR-Ⅱ;chloroform group,ethyl acetate group reduced the content of CTGF、TβR-Ⅱ.Conclusion:(1)Polysaccharide and methanol extract of Dendrobium officinale showed some inhibitory on the proliferation of TGFβ1-induced HSC-T6 cells.(2)N-hexane extract,chloroform extract,ethyl acetate extract had an inhibitory effects on the proliferation of TGFβ1-induced HSC-T6 cells(P<0.05).(2)N-hexane extract is able to inhibit the proliferation of HSC-T6 may be through inhibiting the expression of α-SMA,CTGF,TβR-Ⅱ,while chloroform extract and ethyl acetate extract can inhibit the proliferation of HSC-T6 may be via inhibiting the expression of CTGF,TβR-Ⅱ.