摘要
目的探讨瘦素-p38丝裂原激活蛋白激酶(MAPK)通路在高糖损伤H9c2心肌细胞中的作用。方法应用35 mmol/L葡萄糖处理H9c2心肌细胞以建立高糖损伤细胞模型。应用细胞计数试剂盒检测细胞存活率;双氯荧光素染色荧光显微镜照相测定胞内活性氧水平;Hoechst 33258核染色检测凋亡细胞形态与数量的改变;罗丹明123染色荧光显微镜照相测定线粒体膜电位;Western blot检测瘦素和p38MAPK蛋白的表达水平。结果 35mmol/L葡萄糖处理H9c2心肌细胞明显促进瘦素的表达。在高糖处理H9c2心肌细胞前,应用50μg/L瘦素拮抗剂预处理24 h明显抑制高糖对磷酸化p38MAPK表达的上调作用。瘦素拮抗剂预处理24 h或p38MAPK抑制剂(SB203580)预处理60 min均能抑制高糖对H9c2心肌细胞的损伤作用,使细胞存活率升高,凋亡细胞数量减少,活性氧生成及线粒体膜电位丢失减小。结论瘦素-p38MAPK通路参与高糖对心肌细胞的损伤。
Aim To investigate the role of leptin-p38 mitogen-acrivated protein kinase （MAPK） pathway in high glucose （HG） -induced injury in H9c2 cardiac ceils. Methods H9c2 cardiac cells were treated with HG to establish a model of cellular injury induced by HG. Cell viability was tested by cell counter kit 8 （CCK-8）. The intracellular level of reactive oxygen species （ROS） was measured by dichlorfluorescein staining and photofluorography. The changes in morphology and amount of apoptotic cells were tested by Hoechst 33258 nuclear staining. Mitochondrial membrane poten- tial （MMP） was detected by rhodamine 123 （Rh123） staining followed by photofluorography. The expression levels of leptin and p38MAPK protein were measured by Western blot assay. Results Exposure of H9c2 cardiac cells to 35 mmol/L glucose （HG） markedly enhanced the expression of leptin. Pretreatment of ceils with 50 txg/L leptin antagonist （LA） for 24 h before exposure to HG significantly inhibited HG-induced upregulation of phosphorylated p-p38MAPK ex- pression. Pretreatment of H9c2 cardiac cells with either LA for 24 h or SB203580 （an inhibitor of p38MAPK） for 60 min prior to exposure to HG reduced HG-induced injuries, as evidenced by an increase in cell viability, decreases in apoptotic cells, ROS production and dissipation of MMP. Conclusion The leptin-p38MAPK pathway is involved in HG-in- duced cardiomyocyte injury.
出处
《中国动脉硬化杂志》
CAS
CSCD
北大核心
2014年第5期433-437,共5页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金(H0208)
广东省科技计划项目(2012B031800358)
作者简介
徐文明，硕士，主治医师，研究方向为糖尿病及其并发症的临床与基础，E-mail为xwm920018@163．com。
陈景福，硕士研究生，研究方向为心血管疾病的损伤与保护机制，E-mail为chenjf_gdmc@163．com。
通讯作者 吴铿，主任医师，硕士研究生导师，研究方向为心血管疾病的损伤与保护机制，E-mail：wukeng1245@hotmail．com。
