Effect of medicated serum of Hexuerougan formula on regulates the proliferation and apoptosis of hepatic stellate cells through sphingosine kinase-1
Objective:Investigation of the effect of serum containing Hepatic Soften and Nourish Formula(HXRGF)on the regulation of sphingosine kinase-1(SphK1)in the proliferation and apoptosis of hepatic stellate cells(HSC).Methods:Resistin was applied to treat HSC-T6 cells with Resistin to establish a fibrotic HSC model,and LV-rSphK1-OE and LV-rSphK1-shRNA lentivirus methods to construct stable cell lines with overexpression or low expression of SphK1.The mrna and protein expressions of TGF-β,α-SMA,Col I,FN,Smad and SphK1 were detected by real-time quantitative PCR and Western blot.The cell viability was detected by CCK8 assay and apoptosis was detected by flow cytometry.Results:①Compared with the control group,TGF-β,α-SMA,Col I,FN,SphK1mRNA and protein expression levels were increased in MF-HSC-T6 cells,whil Smade mRNA levels were decreased(all P<0.01).The expression levels of SphK1 and related fibrosis markers were increased in HSC-T6 SphK1 overexpression cell lines(P<0.01).Fibrosis related markers such as TGF-β,α-SMA,Col I and FN mRNA were decreased in HSC-T6 SphK1 low expression cell lines,while Smad mRNA levels were increased(P<0.01).②Compared with model group,the mrna and protein expression levels of MF-HSC T6 SphK1 were decreased after the intervention of TCM compound HXRGF containing serum,along with the expression levels of α-SMA,Col I and FN(P<0.01),and the proliferation rate of HSC cells was decreased(110.85%).Cell proliferation rate decreased and apoptosis rate increased(12.14%)in shRNA-SphK1 transfection group(P<0.01).The apoptosis rate of low expression group of HSC-T6 SphK1 was increased(16.48%,P<0.01).Conclusion:SphK1 can be a potential therapeutic target for hepatic fibrosis.By inhibiting the expression of SphK1,HXRGF can down-regulate the expression level of TGF-β,α-SMA and other pro-fibrosis factors,inhibit the proliferation of activated HSC,induce the apoptosis of HSC,and alleviate liver fibrosis.
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维普
