尼可地尔、谷氨酰胺、美托洛尔单独及合用对大鼠心肌缺血/再灌注损伤后心肌细胞抗凋亡和HSP70的影响  

Effects of nicorandil,glutamine,metoprolol used in single and combination on antiapoptosis ability of cardiocytes and expression of HSP70 against myocardial ischemia /reperfusion injury in rats

作　　者：陈文华[1] 孙畅[1] 张颖[1] 吴艳娜[1] 温克[1] 康毅[1] 娄建石[1] 

机构地区：[1]天津医科大学基础医学院药理学教研室,天津300070

出　　处：《中国药理学通报》2014年第9期1242-1246,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81072631);天津市自然科学基金资助项目(No 09JCZDJC21100)

摘　　要：目的研究尼可地尔(nicorandil,Nic)、谷氨酰胺(glutamine,Glu)、美托洛尔(metoprolol,Met)三药联用对大鼠心肌缺血/再灌注损伤后心肌细胞抗凋亡能力和保护性蛋白HSP70表达的影响。方法将健康♂Wistar大鼠随机分为7组:1假手术(Sham)组,只实施冠状动脉左前降支(left anterior descending,LAD)穿线;2 I/R组,实施LAD 30 min缺血/120 min再灌注程序(ischemia/reperfusion,I/R);3 Nic组;4 Glu组;5 Met组;6三药联用(NGM)组;7三药联用低剂量(NGML)组。药物预处理组则在执行I/R程序前30 min腹腔注射Nic、Glu和Met,三药的初始给药剂量均为1mg·kg-1。TUNEL(terminal dexynucleotidyl transferase-mediated dUTP nick end labeling)法检测心肌细胞凋亡情况,通过凋亡指数衡量心肌细胞凋亡率;Western blot法检测凋亡相关蛋白Bcl-2、Bax以及保护性蛋白热休克蛋白70(heat shock protein 70,HSP 70)的表达情况。结果 I/R组中,缺血区的细胞凋亡率较高(36.9%±10.3%),细胞凋亡相关蛋白表达Bcl-2/Bax值较低(0.14±0.08);与I/R组相比,药物预处理组的细胞凋亡率明显降低(P<0.01),凋亡相关蛋白表达Bcl-2/Bax值明显升高(P<0.01);与Sham组相比,I/R组的HSP70表达明显增加(P<0.01);与I/R组相比,药物预处理组的HSP70表达明显增加(P<0.01);与药物单用组相比,3药联用组的HSP70表达更高(P<0.01)。结论对于大鼠在体I/R损伤,三药联用组与药物单用组相比细胞凋亡率明显降低,凋亡相关蛋白Bcl-2、Bax的表达有所改善;保护性蛋白HSP70的表达增加。Aim To investigate the protective effects of nicorandil, glutamine and metoprolol used in single and combination on the antiapoptosis ability of cardio-cytes and the expression of HSP70 after myocardial is-chemia/reperfusion injury in rats. Methods Male Wistar rats were divided randomly into seven groups：（ 1 ） Sham group：in which the coronary artery was not roped;（2） I/R group：in which only 30 min ischemia and 120 min reperfusion of LAD was executed; （ 3 ） Nicorandil group; （ 4 ） Glutamine group; （ 5 ） Meto-prolol group; （ 6 ） Nic ＋ Glu ＋ Met （ NGM ） group;（7） low dose of Nic＋Glu＋Met （ NGML） group. In the pharmacological precondition groups, correspond-ing drugs were administered 30 min before I/R proto-col, and the initial drug dose in each group was 1 mg ·kg-1 . Myocardial apoptotic rates were measured with TUNEL （ terminal dexynucleotidyl transferase-mediated dUTP nick end labeling） , and the expression of apop-tosis related proteins-Bcl-2/Bax and protective pro-tein-HSP70 was detected by Western blot. ResultsIn I/R group, the apoptotic rate in ischemic region was very high （ 36.9% ± 10.3%） , and Bcl-2/Bax was very low （ 0.14 ±0.08 ） . Compared with I/R group, the apoptotic rate of pharmacological precondition groups was decreased significantly （ P 〈0.01 ） , and the ratio of Bcl-2 to Bax was raised （ P〈0.01 ） . Com-pared with Sham group, the expression of HSP70 in-creased significantly （ P〈0.01 ） in I/R group. Com-pared with IR group, the expression of HSP70 of phar-macological precondition groups was elevated （ P 〈0.01 ） , and the expression in NGM group was higher than in the single drug group （ P〈0.01 ） . Conclusion Compared with single therapy after I/R in rat, the combination therapy of nicorandil, glutamine, meto-prolol can decrease the apoptotic rate, and the expres-sion of apoptosis related proteins is developed. In the mean time, the expression of protective protein HSP70 is elevated.

关 键 词：缺血 再灌注损伤 联合用药 抗凋亡 HSP 70 细胞凋亡率 心脏保护 

分 类 号：R-332[医药卫生] R322.11