H_2O_2预处理对大鼠心肌细胞缺氧/复氧损伤的影响  

Effects of H_2 O_2 Pretreatment on Rat Myocardial Hypoxia/Reoxygenation Injury

作　　者：徐兴华[1] 董福强[1] 田轶魁[1] 魏民新[1] 

机构地区：[1]天津医科大学总医院心血管外科,300052

出　　处：《天津医药》2012年第7期692-694,I0001,共4页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(项目编号:81041042);天津市应用基础及前沿技术研究计划项目(项目编号:10JCZDJC19200);天津医科大学科学基金资助项目(项目编号:2009KY30)

摘　　要：目的:探讨H2O2预处理对大鼠心肌细胞缺氧/复氧引起细胞凋亡的影响。方法:大鼠心肌细胞H9C2经体外培养扩增,使用对数生长期细胞做实验处理。分别用浓度为0、5、10、20、50、100μmol/L的H2O2处理H9C2细胞,以确定最佳H2O2预处理浓度。观察低浓度H2O2预处理对缺氧/复氧所引起细胞损伤的影响。细胞分为对照组、缺氧/复氧组、H2O2预处理组和Janus激酶2抑制剂(AG490)+H2O2组。AnnexinV-FITC/PI双染法及FCM检测细胞凋亡;MTT法检测H9C2细胞增殖活性;WesternBlotting法检测信号转导子和转录激活子3(STAT3)磷酸化水平。结果:20μmol/LH2O2组的STAT3磷酸化水平显著高于其他各浓度组,心肌细胞凋亡率显著低于50及100μmol/L浓度组,心肌细胞活力高于50及100μmol/L浓度组,而与10μmol/L组差异无统计学意义。缺氧损伤后给予AG490处理可使H2O2预处理保护作用消失。结论:20μmol/LH2O2预处理对心肌细胞缺氧/复氧损伤具有适应性保护作用,其可能是通过激活JAK2-STAT3通路发挥抑制细胞凋亡作用的。Objective： To investigate the effect of H2O2 pretreatment on apoptosis caused by hypoxia/reoxygenation in rat cardiac H9C2 ceils. Methods： The H9C2 cells were cultured in vitro for amplification. The logarithmic growth phase cells were used to do the experimental treatment. H9C2 cells were treated with 0, 5, 10, 20, 50 and 100 umol/L concentration of H2O2 respectively, in order to determining the optimal concentration of H2O2 pretreatment. The effect of low concentrations of H2O2 pretreatment on hypoxia/reoxygenation cell injury was observed. Cells were divided into 4 groups, control group, hypoxia/ reoxygenation group, H2O2 pretreatment group and AG490＋H2O2 group. AnnexinV-FITC/PI double staining method and FCM were used to detect apoptosis. MTT assay was used to detect the H9C2 cell proliferation. Western Blotting method was used to detect STAT3 phosphorylation level. Results： The level of STAT3 phosphorylation was higher in H9C2 cell group treated by 20 umol/L H2O2 than that of other groups. The myocardial cell apoptosis rates were significantly lower in H9C2 cell group treated by 20 umol/L H2O2 than those of 50 and 100 umol/L treatment groups. The myocardial cell proliferation was higher in 20 umol/L H2O2 treatment group than that of 50 and 100 umol/L treatment groups, but no significant difference was found compared with 10 umol/L treatment group. The protective effect of H2O2 pretreatment on oxygen injury was abolished after treatment with AG490. Conclusion： Protective effect of 20 umol/L H2O2 pretreatment on myocardial hypoxia/reoxygenation injury was by the activated JAK2-STAT3 pathways to inhibit cell apoptosis.

关 键 词：过氧化氢 再灌注损伤 细胞凋亡 缺血预处理 H9C2细胞 JAK2-STAT3 

分 类 号：R614[医药卫生—麻醉学]