Study On The Function Of MiR-27a In Hepatic Stellate Cells And The Anti-fibrosis Mechanism Of Tanshinone ⅡA

Aim: The aim of this study was to investigate the role of mi R-27 a in hepatic stellate cell(HSC)activation and to explore underlying action mechanism of Tanshinone ⅡA(TIIA)in liver fibrosis.Methods: 1.Real-time quantitative reverse transcription PCR(RT-q PCR)was used to detect the expression of mi R-27a;2.Cell viability assay was used to detect the effect of TIIA and mi R-27 a on HSC proliferation;3.Western Blot experimental method was used to detect the regulation of TIIA and mi R-27 a on hepatic stellate cell activation-related proteins and target genes proteins;4.Network pharmacology approaches were applied to further understand the action of mechanism of TIIA and experimental verification in vivo and in vitro.Results: 1.mi R-27 a was induced by TGF-β1 in LX2 cells and in the culture medium.Inhibition of mi R-27 a attenuates HSCs proliferation(P<0.05)and overexpression of mi R-27 a in activated HSCs upregulated alpha smooth muscle actin(α-SMA)(P<0.05)and Collagen alpha-2(I)(COL1A2)(P<0.01)protein levels.TIIA downregulated mi R-27 a expression(P<0.01)and inhibited α-SMA and COL1A2 protein levels in HSCs(P<0.01).Further studies showed that mi R-27 a mimic treatment reversed TIIA-mediated downregulation of α-SMA and COL1A2 in TGF-β1-treated HSCs(P<0.01).2.TIIA attenuates CCL4-induced liver injury and fibrosis in rats.TIIA inhibited the CCL4-induced serum ALT,AST,ALP and Tbil levels compared with the model group(P<0.05).Additionally,TIIA could decrease Hyp in hepatic level compared with the model group(P<0.01).TIIA significantly decreased the expression of COL1A2 and α-SMA,which were upregulated in the CCL4 model group(P<0.05).3.TIIA significantly downregulated the expression of C-JUN,P-C-JUN,C-MYC,CCND1,MMP9,P65,P-P65,IKBA,PI3 K,CAS9 and P38,which were upregulated during HSC activation in vitro.And TIIA significantly downregulated the expression of C-JUN,C-MYC,MMP9,PI3 K and P38,which were upregulated in the CCL4 model group in vivo.Overexpression of mi R-27 a increased expression of C-JUN,C-MYC and PI3 K protein levels and reversed TIIA-mediated downregulation of C-JUN,C-MYC and PI3 K protein levels in TGF-β1-treated HSCs in vitro.Conclusions: mi R-27 a plays an important role in HSCs activation and can serve as a therapeutic target for hepatic fibrosis.TIIA inhibits,at least in part,HSC proliferation and activation via targeting the mi R-27 a.TIIA could alleviate liver fibrosis through multiple targets and multiple signaling pathways,which provide deep insight into the pharmacological mechanisms of TIIA in the treatment of chronic diseases.