Effect of high fat on fibrosis in rat hepatic stellate cells
10.3760/cma.j.issn.1007-3418.2016.03.007
- VernacularTitle:高脂对大鼠肝星状细胞纤维化的影响
- Author:
Ji YUAN
1
;
Yanyan LIU
;
Xuejiao MA
;
Jiabin LI
Author Information
1. 230022,安徽医科大学第一附属医院感染病科
- Keywords:
Liver cirrhosis;
Hepatic stellate cell;
High mobility group box 1
- From:
Chinese Journal of Hepatology
2016;24(3):191-195
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of high fat on the expression of high-mobility group box 1 (HMGB1),α-smooth muscle actin (α-SMA),and matrix metalloproteinase-2 (MMP-2) in rat hepatic stellate cells (HSC-T6 cells).Methods HSC-T6 cells were cultured in vitro and treated with palmitic acid (PA) at a concentration of 300 μmol/L for 24 hours,and the HSC-T6 cells in the control group were treated with bovine serum albumin (BSA) of the same volume for 24 hours;Westem blot was used to measure the expression of α-SMA and MMP-2.The HSC-T6 cells in the dose-effect group were treated with PA at concentrations of 0,100,200,300,500,and 1000 μmol/L for 24 hours;the HSC-T6 cells in the time-effect group were treated with PA for 0,4,8,12,24,and 48 hours;in the control group,PA was replaced by BSA of the same volume;Western blot was used to measure the expression of HMGB 1,α-SMA,and MMP-2.The HSC-T6 cells were treated with recombinant HMGB1 (rHMGB1) at concentrations of 0,50,100,200,and 500 ng/ml for 48 hours,and in the control group,PA was replaced by phosphate buffer of the same volume.Western blot was used to measure the expression of α-SMA and MMP-2.The HSC-T6 cells in the PA group were treated with PA at a concentration of 300 μmol/L for 24 hours;the HSC-T6 cells in the PA+HMGB1-siRNA group were treated with PA at a concentration of 300 μmol/ L for 24 hours after HMGB1 in HSC-T6 cells was down-regulated;in the blank control group,PA was replaced by BSA of the same volume.Western blot was used to measure the expression ofHMGB1,α-SMA,and MMP-2.One-way analysis of variance was applied for continuous data,and the t-test was applied for comparison between two groups.Results (1) The expression of α-SMA and MMP-2 increased significantly after HSC-T6 cells were treated with PA at a concentration of 300 μmol/L for 24 hours (P ＜ 0.05).(2) Compared with the HSC-T6 cells in the untreated group (0 μmol/L),the HSC-T6 cells treated with different concentrations of PA showed significant increases in the expression of HMGB1 and MMP-2 (P ＜ 0.01),as well as a significant increase in the expression of α-SMA at concentrations of 200,300,500,and 1000 μmol/L (P ＜ 0.01);the HSC-T6 cells treated with PA at a concentration of 300 μmol/L for different periods of time showed varying degrees of increase in the expression of HMGB1,α-SMA,and MMP-2,with significant increases at 16,24,and 48 hours (P ＜ 0.01).(3) Compared with the HSC-T6 cells in the untreated group (0 ng/ml),the HSC-T6 cells treated with rHMGB 1 at concentrations of 100,200,and 500 ng/ml for 48 hours showed significant increases in the expression of α-SMA and MMP-2 (P ＜ 0.05).(4) Compared with the HSC-T6 cells in the PA group,the HSC-T6 cells treated with PA+HMGB1-siRNA for 24 hours showed significant reductions in the expression of HMGB 1,α-SMA,and MMP-2 (P ＜ 0.05).Conclusions High fat can increase the expression of α-SMA and MMP-2 through up-regulating the expression of HMGB 1 in HSC-T6,and thus lead to the development of liver fibrosis.
