饱和氢盐水对心肌缺血再灌注损伤老年大鼠心肌细胞自噬的影响  

Effect of hydrogen-rich saline on cardiomyocyte autophagy during myocardial ischemia-reperfusion in aged rats

作　　者：潘震华[1] 赵悦[1] 于宏颖[1] 刘大一[1] 徐华[1] 

机构地区：[1]大庆油田总医院心脏内科,黑龙江省大庆市163411

出　　处：《中华医学杂志》2015年第25期2022-2026,共5页National Medical Journal of China

摘　　要：目的 探讨饱和氢盐水对心肌缺血再灌注损伤老年大鼠心肌细胞自噬的影响.方法 选取健康清洁级雄性SD大鼠150只,18月龄,体重400 ～ 540 g.采用随机数字表法将其分为5组（n=30）：空白对照组（Ⅰ组）、假手术组（Ⅱ组）、缺血再灌注组（Ⅲ组）、饱和氢盐水治疗组（Ⅳ组）、生理盐水治疗组（Ⅴ组）.Ⅰ组不做任何处理;Ⅱ组大鼠仅穿线不结扎;Ⅲ组制备大鼠心肌缺血再灌注损伤模型;Ⅳ组于再灌注前5 min给予1 ml/100 g腹腔注射饱和氢生理盐水;Ⅴ组于再灌注前5 min给予1 ml/100 g腹腔注射等量生理盐水.采用HE染色观察心肌病理学,采用MDC荧光染色,流式细胞仪定量检测心肌细胞自噬率,采用蛋白质印迹法检测心肌细胞腺苷酸活化蛋白激酶（AMPK）、雷帕霉素靶蛋白（mTOR）、Beclin基因1（Beclin1）、微管相关蛋白1轻链3（LC3）蛋白含量.结果 与Ⅰ组、Ⅱ组比较,Ⅲ组、Ⅳ组和Ⅴ组大鼠心肌缺血再灌注后12、24 h心肌细胞自噬率升高（F=23.45,26.65,25.58）,心肌细胞AMPK（F=23.16,25.15,27.85）、mTOR（F=21.04,24.83,27.43）、Beclin1（F =22.15,25.79,29.05）和LC3（F =22.58,27.25,28.46）蛋白表达升高（均P＜0.05）.与Ⅲ组、Ⅴ组比较,Ⅳ组大鼠心肌缺血再灌注后12、24h心肌细胞自噬率降低（F=21.29,24.71）,心肌细胞AMPK（F=22.37,25.84）、mTOR（F=20.48,22.38）、Bedin1 （F=21.76,28.43）和LC3（F=22.54,27.21）蛋白表达降低（均P＜0.05）.结论 饱和氢盐水可通过降低心肌细胞自噬,减轻老年大鼠心肌缺血再灌注损伤,其机制可能与降低心肌细胞AMPK、mTOR、Beclin1和LC3蛋白表达有关.Objective To investigate the effects of hydrogen-rich saline on cardiomyocyte autophagy during myocardial ischemia-reperfusion in aged rats.Methods One hundred and fifty healthy male Sprague Dawley rats,18 months old,weighing 400-540 g were selected.The rats were then randomly divided into 5 groups（n =30）：Normal control group（group Ⅰ）;Sham operation group（group Ⅱ）;Myocardial ischemiareperfusion group （group Ⅲ）;Hydrogen-rich saline group（group Ⅳ）;Normal saline group（group Ⅴ）.No any processing in group Ⅰ.In group Ⅱ,the anterior descending branch was only exposed but not ligated.Myocardial I/R was induced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 12 h and 24 h of reperfusion with Bimbaum.Hydrogen-rich saline 1 ml/100 g were injected intraperitoneally 5 min before reperfusion in group Ⅳ.Normal saline 1 ml/100 g were injected intraperitoneally 5 min before reperfusion in group Ⅴ.The rats were sacrificed at 12 h and 24 h of reperfusion and hearts were removed.The pathological changes of myocardial tissue were detected by HE staining.The rate of cardiomyocyte autophagy were detected by the MDC fluorescent dye and flow cytometry instrument.The expression of AMPK,mTOR,Beclin1,LC3 in myocardial tissue was investigated by Western blot.Result Compared with groups Ⅰ and Ⅱ,the rate of cardiomyocyte autophagy,the expression of AMPK,mTOR,Beclin1,LC3 in myocardial tissue were significantly increased at 12 h,24 h in groups Ⅲ,ⅣandⅤ （F =23.45,26.65,25.58;F =23.16,25.15,27.85;F =21.04,24.83,27.43;F =22.15,25.79,29.05;F=22.58,27.25,28.46）,P 〈0.05.Compared with group Ⅲ and Ⅴ,the rate of cardiomyocyte autophagy,the expression of AMPK,mTOR,Beclin1,LC3 were significantly decreased at 12 h,24 h in group Ⅳ（F=21.29,24.71;F=22.37,25.84;F=20.48,22.38;F=21.76,28.43;F=22.54,27.21）,P〈 0.05.Conclusion Hydrogen-rich saline can attenuate myocardial reperfusion injury through inhibiting cardiomyocyte autophagy.The mechanism may be a

关 键 词：心肌缺血 再灌注损伤 老年人 氢 自噬 

分 类 号：R541.4[医药卫生—心血管疾病]