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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 establish an in vitro model of ethanol-induced hepatocellular injury and to investigate the expression and significance of nuclear factor NF-κB1 (p65) in ethanol-induced hepatocellular injury. Methods Human normal hepatocyte L02 strain was cultured in vitro. The hepatocyte injury was induced by different concentrations of ethanol. The content of aminotransferase in the supernatant of the cells was detected. The expression and localization of NF-κB 1 protein were observed by immunohistochemistry. NF-κB1 mRNA expression level. Results Compared with the control group, the expression of aspartate aminotransferase (AST) in 40, 60, 80 and 100 mmol / L ethanol groups was significantly higher than that in the control group (P <0.01) (P <0.05, P <0.01); Immunocytochemistry With the increase of ethanol dose, the expression of NF-κB in the cytoplasm nucleus gradually (P <0.01). The expression of NF-κB mRNA in 40,60,80,100 mmol / L group was higher than that in control group by real-time quantitative PCR (P <0.01). Conclusion NF-κB1 is involved in the development of alcoholic hepatocyte injury in vitro.