NF-κB1在体外乙醇诱导损伤肝细胞中的表达及意义

Expression of Nuclear Factor-kappa B in Alcohol-induced Impaired Hepatocytes and its Significance

目的建立乙醇诱导肝细胞损伤的体外模型,探讨核因子NF-κB1(p65)在乙醇诱导肝细胞损伤中的表达及意义。方法体外培养人正常肝细胞L02株,以不同浓度乙醇诱导肝细胞损伤,检测细胞上清液中转氨酶的含量,免疫组织化学法观察细胞中NF-κB1蛋白的表达强度及定位,实时定量PCR检测NF-κB1mRNA表达水平。结果乙醇染毒40、60、80、100mmol/L组天门冬氨酸氨基转移酶(AST)表达量高于对照组,差异有统计学意义(P<0.01);80、100 mmol/L组谷氨酸转氨酶(GGT)的表达量高于对照组,差异有统计学意义(P<0.05,P<0.01);免疫细胞化学随着乙醇剂量增加,胞浆内胞核内NF-κB1的表达逐渐增加;实时定量PCR检测40、60、80、100 mmol/L组NF-κB1mRNA表达量高于对照组,差异有统计学意义(P<0.01)。结论 NF-κB1参与了体外酒精性肝细胞损伤的发展过程。

Objective To investigate the expression and its significance of nuclear factor-kappa B1 （NF-κB1, p65） in alcohol-induced impaired hepatocytes. Methods Alcohol induces human hepatocytes L02 injury in different concentrations, the quantity of transaminase were determined. NF-κB1 protein was observed under light microscope by immunohistochemistry. The expression of NF-κB1 mRNA was determined by real time-PCR. Results After L02 cells exposure to alcohol in 40, 60, 80, 100mmol/L, respectively, the contents of aspartate transaminase （AST） in these groups were significantly increased （P〈0.01）. After exposure to alcohol in 80 and 100mmol/L dose, the contents of glutamate transaminase （GGT） were significantly increased （P〈0.05, P〈0.01）. The expressions of NF-κB1 protein in the alcohol treated groups were higher than that in the control group. The levels of NF-κB1 mRNA expression in the 40, 60, 80, 100mmol/L alcohol treated groups were significantly higher than that in the control group（P〈0.Ol）. Conclusion NF-κB1 may be involved in the development of the alcohol-induced hepatocytes injury in vitro.