活性氧与丝裂原激活蛋白激酶通路的相互作用介导高糖引起的心肌细胞损伤

目的探讨活性氧(ROS)与丝裂原激活蛋白激酶(MAPK)通路的相互作用在高糖损伤H9c2心肌细胞中的作用。方法应用细胞计数盒(CCK-8)检测细胞存活率;Hoechst 33258核染色检测凋亡细胞形态及数量的改变;双氯荧光素(DCFH-DA)染色荧光显微镜照相检测细胞内ROS水平;Western blot测定蛋白质表达水平。结果高糖(35 mmol/L葡萄糖)处理H9c2心肌细胞24 h可引起明显的损伤,表现为细胞存活率下降,凋亡细胞数量及ROS水平明显升高。另方面,高糖可明显地上调磷酸化(p)p38MAPK、细胞外信号调节蛋白激酶1/2(ERK1/2)及c-Jun N端激酶(JNK)(为MAPK家族的3个成员)的表达水平。N-乙酰半胱氨酸(NAC,为ROS清除剂)能抑制高糖引起的心肌细胞毒性和细胞凋亡,也能阻断高糖对p-p38MAPK、p-ERK1/2及p-JNK表达的上调作用。此外,p38MAPK、ERK1/2和JNK的选择性抑制剂均能抑制高糖引起的心肌损伤,并能抑制ROS生成增多。结论在高糖损伤H9c2心肌细胞中,存在ROS与MAPK通路的正相互作用,这种相互作用可能在高糖引起的心肌细胞损伤中起着重要的作用。

Interaction between ROS and MAPK pathway mediates high glucose-induced injury in H9c2 cardiac cells

Objective To explore the role of interaction between reactive oxygen species(ROS) and mitogen-activated protein kinase(MAPK) pathway in high glucose(HG)-induced injury in H9c2 cardiac cells.Methods Cell viability was measured by cell counter kit(CCK-8);The changes in morphology and amount of apoptotic cells were tested by Hoechst nuclear staining.The level of intracellular ROS was detected by DCFH-DA staining and photofluorography.The expression level of MAPK protein was tested by Western blot assay.Results Exposure of H9c2 cells to HG(35 mmol / L glucose) for 24 h induced significant injuries,as evidenced by a decrease in cell viability,increases in apoptotic cell amount and ROS production.On the other hand,HG markedly upregulated phosphorylated(p) p38MAPK,p-extracellular signal-regulated protein kinase 1 / 2(ERK1 / 2) and c-Jun Nterminal kinase(JNK)(three members of MAPK) expression level.N-acetyl-cystein(NAC,a ROS scavenger) inhibited HG-induced cytotoxicity and cell apoptosis as well as upregulation of p-p38MAPK,p-ERK1 / 2 and p-JNK expression.Furthermore,the selective inhibitors of p38MAPK,ERK1 / 2 and JNK reduced not only HG-induced cardiomyocyte injury,but also overproduction of ROS.Conclusion During H9c2 cells injury induced by HG,there is a positive interaction between ROS and MAPK pathway.This interaction may play an important role in HG-induced injury in H9c2 cells.