p38 MAPK在高糖高脂诱导的心肌细胞损伤及氧化应激中的作用研究  

The role of p38 MAPK in the high glucose and fat induced cardiomyocyte injury and oxidative stress

作　　者：杜发旺[1] 王纪人[1] 刘兴会[1] 潘婷婷[1] 刘会[1] 谭洪文 陈保林[1] DU Fawang;WANG Jiren;LIU Xinghui;PAN Tingting;LIU Hui;TAN Hongwen;CHEN Baolin(Department of Cardiology,Guizhou Provincial People′s Hospital,Guiyang,Guizhou 550002,China)

机构地区：[1]贵州省人民医院心内科,贵州贵阳550002

出　　处：《现代医药卫生》2023年第12期1981-1986,1992,共7页Journal of Modern Medicine & Health

基　　金：贵州省卫生健康委科学技术基金项目(gzwjkj2019-1-097);贵州省科学技术厅基础研究项目(黔科合基础[2019]1199);贵州省科学技术厅临床研究中心项目(黔科合平台人才[2017]5405);贵州省科技厅临床专项黔科合成果(LC[2021]027);贵州省人民医院青年基金项目(GZSYQN[2019]10号)。

摘　　要：目的 探讨p38丝裂原活化蛋白激酶(p38 MAPK)在高糖高脂诱导的心肌细胞损伤及氧化应激中的作用。方法 高糖(33 mmol/L)及不同浓度高脂(棕榈酸钠125、250、500、1 000、2 000、4 000μmol/L)不同时间(12、24、48、72 h)干预H9c2心肌细胞,选择合适的棕榈酸钠浓度及干预时间。后续将H9c2心肌细胞分为对照组(H9c2组)及高糖高脂组,应用乳酸脱氢酶(LDH)试剂盒检测细胞培养液中LDH的活性,采用实时荧光定量聚合酶链反应(RT-PCR)检测心肌细胞肥大指标心房钠尿肽(ANP)、α-心肌肌动蛋白(α-SKA)mRNA的表达,通过western blotting技术检测高糖高脂干预H9c2心肌细胞72 h p38磷酸化/GAPDH的蛋白表达量,p38 MAPK的特异性抑制剂(SB203580)分别测定各组中活性氧(ROS)、一氧化氮(NO)的水平。结果 相比对照组,高糖高脂组LDH的释放率明显升高,差异有统计学意义(P<0.05)。进一步用TUNEL法检测细胞凋亡,结果显示,相比对照组,高糖高脂组细胞凋亡指数也明显升高,差异有统计学意义(P<0.05)。高糖高脂可增加H9c2细胞中ANP及α-SKA mRNA水平的表达,相比对照组,其H9c2细胞中ANP及α-SKA mRNA水平明显升高,差异有统计学意义(P<0.05)。与对照组相比,高糖高脂组中p38磷酸化水平明显升高,差异有统计学意义(P<0.05)。高糖高脂组中的NO水平远远高于对照组,而SB203580组中NO水平明显低于高糖高脂组,差异均有统计学意义(P<0.05)。同样,高糖高脂组中的ROS水平明显高于对照组,而SB203580组中NO水平明显低于高糖高脂组,差异均有统计学意义(P<0.05)。结论 p38 MAPK通路参与高糖高脂诱导的心肌细胞损伤及氧化应激,通过抑制p38 MAPK表达可以抑制高糖高脂诱导的氧化应激反应。Objective To investigate the role of p38 mitogen activated protein kinase(p38 MAPK)in cardiomyocyte injury and oxidative stress caused by high glucose and fat levels.Methods At different times(12,24,48,72 h),high glucose(33 mmol/L)and high fat concentrations(125,250,500,1000,2000,4000μmol/L)were used to intervene the H9c2 cardiomyocytes,so as to select appropriate sodium palmitate concentrations and intervention time.In the following experiment,H9c2 cardiomyocytes were divided into two groups:the control group(the H9c2 group),and high glucose and high fat group.Lactate dehydrogenase(LDH)kit was used to detect LDH activity in cell culture medium,the mRNA expression of hypertrophic indicators including atrial natriuretic peptide(ANP)andα-myocardial actin(α-SKA)were detected by fluorescence real-time quantitative polymerase chain reaction(RT-PCR),and the protein expression of p-p38/GAPDH in H9c2 cardiomyocytes treated with high glucose and fat was detected by western blotting technique at 72 h.The specific inhibitor of p38 MAPK(SB203580)was used to determine the levels of reactive oxygen species(ROS)and nitric oxide(NO)in each group.Results In the high glucose and high fat group,the release rate of LDH was significantly higher than in the control group(P<0.05).In addition to detecting apoptosis by TUNEL,the results showed the apoptosis index of the high glucose and high fat group was significantly higher than that of the control group(P<0.05).High glucose and fat could increase the expression of ANP andα-SKA mRNA levels in H9c2 cells,relative to the control group,the ANP andα-SKA mRNA levels in H9c2 cells were significantly increased,and the differences were statistically significant(P<0.05).Compared with the control group,the phosphorylation level of p38 in the high glucose and high fat group was significantly increased,and and there was a statistically significant difference(P<0.05).The level of NO in the high glucose and high fat group was significantly higher than that in the control group,while the level o

关 键 词：丝裂原活化蛋白激酶 高糖高脂 糖尿病心肌病 氧化应激 调控作用 

分 类 号：R363[医药卫生—病理学] R34[医药卫生—基础医学]