Augmenter of liver regeneration promotes the proliferation of HL-7702 cells in carbon tetrachloride-induced acute liver injury via increasing autophagy
10.3760/cma.j.issn.1007-3418.2016.10.009
- VernacularTitle:肝再生增强因子通过增加自噬水平促进CCl4诱导的急性肝损伤中HL-7702细胞的增殖
- Author:
Weijia HAN
1
;
Hongbo SHI
;
Honglin SHI
;
Jinyue SONG
;
Feng REN
;
Zhongping DUAN
;
Yu CHEN
Author Information
1. 100069,首都医科大学附属北京佑安医院人工肝中心
- Keywords:
Liver regeneration;
Liver injury,acute;
Augmenter of liver regeneration;
Autophagy
- From:
Chinese Journal of Hepatology
2016;24(10):761-766
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the protective effect of augmenter of liver regeneration (ALR) against acute liver injury and related mechanisms.Methods HL-7702 cells were divided into normal control group,carbon tetrachloride (CCl4)-induced acute liver injury group,ALR+CCl4 intervention group,3-methyladerine (3-MA)+CCl4 intervention group,and ALR+3-MA+CCl4 intervention group.The ALR+CCl4 and ALR+3-MA+CCl4 intervention groups were transfected with ALR plasmids at 8 hours before CCl4 treatment.All groups except the normal control group were treated with CC14,and 30 minutes later,the 3-MA+CC14 and ALR+3-MA+CCl4 intervention groups were treated with 3-MA.The cells were collected at 24 hours after CCl4 treatment.The HL-7702 cells and supematant were collected to measure the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) (IU/L).Westem blot was used to measure the levels ofALR,cyclin D,cyclin E,proliferating cell nuclear antigen (PCNA),autophagy-related gene 7 (Atg7),and autophagy genes LC3,p62,and Beclin-1.Quantitative real-time PCR was used to measure the mRNA expression ofALR.A oneway analysis of variance was used for comparison of means between any two groups.Results The ALR+CCl4 intervention group had significant increases in the protein and mRNA expression of ALR compared with the acute liver injury group (both P ＜ 0.05).The CC14-induced acute liver injury group had significant increases in the protein and mRNA expression of ALR compared with the normal control group (both P ＜ 0.05).Compared with the CCl4-induced acute liver injury group,the ALR+CCl4 intervention group had significant reductions in ALT (0.73±0.17 IU/L vs 1.43±0.38 IU/L,P ＜ 0.05) and AST (19.85±1.83 IU/L vs 56.73±6.25 IU/L,P ＜ 0.05) in supematant,significantly increased expression of cyclin D,cyclin E,PCNA,LC3,Atg7,and Beclin-1 in hepatocytes,and significantly reduced expression of p62,which suggested that ALR protected the liver against acute liver injury,promoted the regeneration of hepatocytes,and enhanced the autophagy of hepatocytes.The ALR+3-MA+CCl4 intervention group had a significant reduction in the expression of regeneration-associated proteins compared with the ALR+CCl4 intervention group,while there was no significant difference between the ALR+3-MA+CCL4 intervention group and 3-MA+CCL4 intervention group,which suggested that after the inhibition of autophagy,there were significant reductions in the regeneration of hepatocytes and liver regeneration promoted by ALR.Conclusion ALR can promote the regeneration of hepatocytes in liver parenchyma,which is achieved by the regulation of autophagy.
