Resveratrol-Pretreated Adipose-Derived Mesenchymal Stem Cells Protect CCl4-Induced Hepatocyte Injury by Regulating Autophagy
LIU Shuangjuan#, SUN Yingying#, HOU Leyao, MA Xiyuan, LÜ Qiongxia, CHEN Xiaoguang, LIU Yumei, ZHANG Ziqiang*
The aim of this study was to investigate the therapeutic effect and mechanism of Res (resveratrol) pretreatment of ADSCs (adipose mesenchymal stem cells) on liver cell injury induced by CCl4 (carbon tetrachloride). ADSCs were isolated and extracted from the groin of mice, cultured to the third generation, and the ability of adipogenic and osteogenic differentiation was detected. The third-generation ADSCs pretreated with different concentrations of Res were co-cultured with CCl4-injured BRL-3A cells. The viability of hepatocytes was detected by CCK-8 method, and the contents of AST and ALT in BRL-3A cells were detected by kit to screen the optimal concentration of Res. Then BRL-3A cells were divided into control group, CCl4 model group, ADSCs treatment group, Res pretreatment ADSCs treatment group, Res pretreatment ADSCs+3-MA treatment group. The contents of AST, ALT, ALB and ALP in each group were detected by kit. Western blot was used to detect the expression of autophagy-related proteins Beclin-1, LC3II, p62 and PI3K/AKT/mTOR pathway-related proteins. The results showed that the optimal concentration of Res was 20 μmol/L. Compared with the model group, ADSCs and Res pretreated ADSCs improved the levels of AST, ALT, ALP and ALB in damaged BRL-3A cells, and the treatment effect of Res pretreatment group was more obvious. In addition, Western blot results showed that compared with the model group, Res-pretreated ADSCs could up-regulate the expression of autophagy-related proteins Beclin-1 and LC3II in BRL-3A cells, down-regulate the expression of p62, and inhibit the expression of AKT/PI3K/mTOR pathway-related proteins. When the autophagy inhibitor 3-MA was added, the therapeutic effect of Res-pretreated ADSCs on injured hepatocytes was significantly inhibited. These results indicate that Res pretreated ADSCs can repair liver injury by promoting autophagy, and the increase of autophagy may be related to the inhibition of PI3K/AKT/mTOR pathway.
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