Effects of 3-methyladenine on activation and autophagy of rat hepatic stellate cell line HSC-T6 induced by TGF-β
Objective To investigate the effects of 3-methyladenine(3-MA)on activation and autophagy of rat he-patic stellate cell line HSC-T6 induced by transforming growth faction-β(TGF-β).Methods HSC-T6 cells in the loga-rithmic growth phase were divided into the blank group,TGF-β+PBS group,and TGF-β+3-MA group.Cells of TGF-β+3-MA group were cultured with 2 ng/mL TGF-β for 72 h and treated with 3-MA(0.5 mg/mL)for 24 h.Cells of TGF-β+PBS group were cultured with 2 ng/mL TGF-β for 72 h and treated with the same volume of PBS for 24 h.Cells in the blank group were treated with equal volume of PBS,and the mRNA levels of cell activation markers α-SMA and TGF-β and au-tophagy markers LC3,Beclin-1 and Atg5 in each group were detected by real-time fluorescence quantitative PCR.Cell ac-tivation markers α-SMA,TGF-β proteins and autophagy markers LC3,Beclin-1,Atg5 proteins were detected by Western blotting.Results The mRNA and protein levels of cell activation markers α-SMA and TGF-β in the TGF-β+PBS group and TGF-β+3-MA group were higher than those in the blank group,and those were lower in the TGF-β+3-MA group than in the TGF-β+PBS group(all P<0.05).The mRNA and protein levels of autophagy markers LC3,Beclin-1 and Atg5 in the TGF-β+PBS group and TGF-β+3-MA group were higher than those in the blank group,and those were lower in the TGF-β+3-MA group than in the TGF-β+PBS group(all P<0.05).Conclusion 3-MA can inhibit TGF-β-induced HSC-T6 activation and autophagy in rat hepatic astrocytes.
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