[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).


CONCLUSIONS
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.
Department of Gastroenterology, Jiangxi Key Laboratory of Molecular Medicine, Nanchang University, Nanchang, China.
Man
M
Liu
Yue
Y
He
Jixiang
J
Zhang
D000199:Actins / Q000235:genetics / Q000378:metabolism; D000225:Adenine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D001343:Autophagy / Q000187:drug effects* / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D016195:G2 Phase / Q000187:drug effects / Q000235:genetics; D055166:Hepatic Stellate Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D000199:Actins; D000225:Adenine; D024042:Collagen Type I; D008869:Microtubule-Associated Proteins; C526109:Anxa3 protein, rat; C498862:Acta2 protein, rat; C025946:3-methyladenine