[Effect of autophagy inhibitor 3-methyladenine on proliferation and activation of hepatic stellate cells].

OBJECTIVE

To investigate the effect of autophagy inhibitor 3-methyladenine (3-MA) on proliferation and activation of hepatic stellate cells (HSCs) and investigate the underlying molecular mechanism.

METHODS

Cultured HSC-T6 cells were cultured in vitro and treated with different concentrations of 3-MA (low-dose group, 2.5 mmol/L; middle-dose group, 5 mmol/L; high-dose group, 10 mmol/L; control group, 0 mmol/L). The mRNA expressions of α-SMA and type I collagen were determined by RT-PCR; the protein expressions of LC3II, a-SMA and type I collagen were detected by Western blotting; cell proliferation was observed by MTT assay and cell cycle by flow cytometry.

RESULTS

The autophagy of HSC-T6 cells decreased with the rising of the concentration of 3-MA. The mRNA expressions of α-SMA and type I collagen in all three 3-MA-treated groups were significantly down-regulated, compared with those in control group (P<0.05). Meanwhile, compared with the control group, the 3-MA-treated groups also showed significantly down-regulated protein expressions of LC3II, α-SMA, and type I collagen (P<0.05), significantly lower proliferation activity (P<0.05) and significantly higher numbers of HSCs in the G2 phase (P<0.05).

CONCLUSION

The autophagy inhibitor 3-MA significantly down-regulated the expressions of LC3II, α-SMA and type I collagen in HSC-T6 and caused an arrest in the G2 phase of the cells, thus inhibiting the proliferation and activation of HSCs.