目的研究并探讨JNK活性对胰岛素信号传递的影响。方法用不同种类的JNK抑制剂处理人肝癌细胞株(HepG2)细胞12h,随后加入10nmol/L胰岛素孵育5min以激活细胞胰岛素信号通路,收集细胞蛋白样品用Western印迹方法检测丝裂原活化蛋白激酶(MAPK)及胰岛素信号通路分子蛋白表达情况。结果JNK抑制剂(JNKi-Ⅷ)可有效抑制HepG2细胞中JNK的活性,并且JNKi—Ⅷ可浓度依赖性地抑制胰岛素信号传递。与JNK-Ⅷ类似,其他JNK抑制剂(JNKi—V、JNKi—Ⅲ和sP600125)均可抑制HepG2细胞中的胰岛素信号通路。结论在肝实质细胞HepG2中,抑制JNK活性可阻断胰岛素信号通路。
Objective To investigate the effects of JNK inhibition on insulin signaling pathway. Methods HepG2 cells were treated with different kinds of JNK inhibitors for 12 h, and then the cells were treated with 10 nmol/L insulin for 5 min to stimulate insulin signaling pathway. Mitogen-activated protein kinases ( MAPK ) and insulin signaling pathways were analyzed by Western blot using the total cell lysates. Results JNK activity was significantly inhibited by JNK inhibitor JNKi-Ⅷ and results showed that JNKi-Ⅷ treatment could reduce insulin signaling pathway in a dose-dependent manner. Furthermore, other JNK inhibitors including JNKi-Ⅴ, JNKi-Ⅲ, and SP600125 blocked JNK activity in HepG2 cells. Similar to JNKi-Ⅷ, these JNK inhibitors also impaired insulin signaling transduction in a dose-dependent manner. Conclusion In HepG2 hepatocytes, JNK activity inhibition blocks insulin signaling transduction.