C-Jun氨基末端激酶与Activin A/Smads通路在体外脑缺血损伤中的相互作用

Interactions between c-Jun N-terminal kinase and Activin A/Smads signal in cerebral ischemic injury in vitro

目的探讨c-Jun氨基末端激酶(JNK)与Activin A/Smads信号在体外缺血性脑损伤中的相互作用机制。方法使用鼠神经生长因子诱导大鼠嗜铬细胞瘤PC12细胞神经元样转化,利用连二亚硫酸钠(NaS2O4)构建细胞氧糖剥夺模型(OGD),计时0h和1h。Western blot检测外源性ActA或JNK磷酸化抑制剂SP600125干预后,OGD不同时间Smad3、JNK1总蛋白和磷酸化蛋白的表达情况。结果 OGD损伤后JNK1磷酸化蛋白表达升高27.1%。与对照组相比,外源性ActA下调JNK1蛋白的磷酸化水平,在OGD 0h和1h时分别下降了67.8%和56.4%。SP600125通过抑制JNK1磷酸化,上调Smad3的磷酸化水平。与DMSO对照组相比,在OGD 0h和1h时,SP600125组磷酸化Smad3表达分别升高了142.5%和60.5%。结论 JNK与ActA/Smads通路之间存在负性调控作用。

Objective To explore the interactions between c-Jun N-terminal kinase(JNK)and Activin A/Smads signal in cerebral ischemic injury in vitro.Methods With the use of nerve growth factor,rattus pheochromocytoma PC12 cell was transferred to nerve-like cell.Then they were exposed to oxygen glucose deprivation(OGD)injury to simulate cerebral ischemia in vitro.With the introduction of exogenous Activin A(ActA)and JNK inhibitor(SP600125)before OGD treatment,the expression changes of total and phosphorylated Smad3 and JNK1 were detected by Western blot.Results After OGD injury,the expressions of phosphorylated JNK1 increased by 27.1%.Act A administration suppressed JNK phosphorylation.At OGD 0 hand 1 h,it decreased by 67.8%and 56.4%respectively.In addition,SP600125 could elevate the phosphorylation of Smad3 through JNK inhibition.At OGD 0 hand 1 h,the level of phosphorylated Smad3 in SP600125 group increased by 142.5%and 60.5%,compared to that in DMSO group.Conclusion There was a negative regulation effect between JNK and ActA/Smads signaling in ischemic injury.