p38MAPK抑制剂联合辅酶Q10对氧化应激条件下大鼠心肌细胞的保护作用  

Protective effects of p38MAPK inhibitor combined with coenzyme Q10 on myocardial cells under oxidative stress

作　　者：张思洁[1] 吴亚琼 李星涛[1] ZHANG Sijie;WU Yaqiong;LI Xingtao(Department of Cardiology,the Fourth Hospital,Hebei Medical University,Shijiazhuang 050000)

机构地区：[1]河北医科大学第四医院心内科

出　　处：《郑州大学学报（医学版）》2019年第5期699-703,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：河北医学科学研究重点课题计划项目(20190707)

摘　　要：目的:研究p38丝裂原活化蛋白激酶(p38MAPK)抑制剂SB203580联合辅酶Q10对氧化应激条件下大鼠心肌细胞H9c2的保护作用。方法:H9c2细胞分为5组,对照组常规培养,模型组给予100μmol/L过氧化氢处理,辅酶Q10组给予100μmol/L的过氧化氢、0.3 g/L辅酶Q10处理,SB203580组给予100μmol/L的过氧化氢、10μmol/L SB203580处理,辅酶Q10+SB203580组给予100μmol/L的过氧化氢、0.3 g/L辅酶Q10、10μmol/L SB203580处理。24 h后,采用Western blot法检测细胞中p38MAPK磷酸化水平,二硝基苯肼显色法检测细胞乳酸脱氢酶(LDH)漏出率,MTT法检测存活率,Annexin V-FITC和PI双染法检测细胞凋亡,Western blot法检测细胞中活化的Caspase-3(C-caspase-3)蛋白水平,硫代巴比妥酸法检测丙二醛(MDA)含量,黄嘌呤氧化法检测超氧化物歧化酶(SOD)活性,钼酸铵法检测过氧化氢酶(CAT)活性。结果:辅酶Q10和SB203580均可抑制氧化应激条件下心肌细胞中p38MAPK磷酸化水平,提高心肌细胞存活率,降低LDH漏出率,降低细胞凋亡率,下调细胞中C-caspase-3蛋白表达水平,提高细胞中SOD和CAT活性,减少细胞中MDA含量,并且二者联用保护作用更强(P均<0.05)。结论:p38MAPK抑制剂联合辅酶Q10可抑制氧化应激诱导的心肌细胞损伤。Aim:To discuss the effects of p38MAPK inhibitor SB203580 combined with coenzyme Q10 on myocardial H9c2 cells under oxidative stress.Methods:H9c2 cells were allocated into 5 groups,control group(conventionally cultured),model group(treated with 100μmol/L H 2O 2),coenzyme Q10 group(treated with 100μmol/L H 2O 2 and 0.3 g/L coenzyme Q10),SB203580 group(treated with 100μmol/L H 2O 2 and 10μmol/L SB203580),and coenzyme Q10+SB203580 group(treated with 100μmol/L H 2O 2,0.3 g/L coenzyme Q10 and 10μmol/L SB203580).After 24 hours,the level of p38MAPK phosphorylation in cells was detected by Western blot,the leakage rate of LDH was detected by dinitrophenylhydrazine colorimetry,the survival rate was detected by MTT assay,apoptosis was detected by Annexin V-FITC and PI double staining,Western blot was used to detect the level of C-caspase-3 protein in cells,the content of MDA was detected by thiobarbituric acid,the activities of SOD and CAT were detected by xanthine oxidation and ammonium molybdate method.Results:Coenzyme Q10 and SB203580 could both inhibit p38MAPK phosphorylation in H9c2 cells under oxidative stress,increase the survival rate,reduce LDH leakage rate and apoptosis rate,down-regulate the expression level of C-caspase-3 protein,increase the activities of SOD and CAT,and reduce MDA content in cells(P<0.05).Moreover,coenzyme Q10 combined with SB203580 had better protective effects on H9c2 cells under oxidative stress(P<0.05).Conclusion:p38MAPK inhibitor combined with coenzyme Q10 could inhibit oxidative stress-induced cardiomyocyte injury.

关 键 词：心肌细胞 辅酶Q10 P38丝裂原活化蛋白激酶 氧化应激 

分 类 号：R541[医药卫生—心血管疾病]