白细胞介素1β对大鼠肝细胞毒性作用的细胞内信号传导途径

Intracellular signal transduction pathways of IL-1β-induced cytotoxicity on primary cultured rat hepatocytes

目的 研究白细胞介素 1β(IL 1β)对原代培养大鼠肝细胞细胞毒性作用的细胞内信号传导机制。 方法 使用雄性Wistar大鼠 ,原位胶原酶灌注分离肝细胞。应用比色法测定乳酸脱氢酶 (LDH)活性 ,WesternBlot方法分析c JunN末端激酶 (JNK)、p38激酶的表达 ,凝胶电泳移动抑制实验检测激活物蛋白 1(AP 1)的结合活性。 结果 IL 1β促进原代培养大鼠肝细胞LDH释放 (IL 1β刺激组与对照组LDH活性分别为 2 1 9%± 3 6 %和 11 0 %± 1 8% ,P <0 0 1) ;IL 1β通过激活JNK途径 ,激活转录因子AP 1,对原代培养大鼠肝细胞产生细胞毒性作用 ,而同时激活的p38激酶途径对这一过程起负性调节作用。 结论 IL 1β通过激活JNK途径 ,激活转录因子AP 1。

Objective To investigate the intracellular signal transduction pathways of IL 1β induced cytotoxicity on primary cultured rat hepatocytes. Methods All studies were based on primary cultured hepatocytes from male Wistar strain rats (5 to 7 weeks old, 170 to 230 g). The activity of lactose dehydrogenase (LDH) was detected by colormetric assay. JNK and p38 kinase were analyzed by Western blot assay. The AP 1 activity was evaluated by electrophoretic mobility shift assay. Results IL 1β significantly increased LDH release. LDH activity in group IL 1β and controls was 21 9%±3 6% and 11 0%±1 8% respectively, P <0 01). IL 1β induced cytotoxicity via activating JNK pathway and transcription factor AP 1 and the simultaneous activation of p38 kinase pathway negatively regulated this process. Conclusion IL 1β induces cytotoxicity in primary cultured rat hepatocytes via activating JNK pathway and transcription factor AP 1.