Chymase抑制对糖尿病大鼠心肌细胞凋亡的影响  

Effect of Chymase Inhibition on Cardiomyocyte Apoptosis in Diabetic Rats

作　　者：王思宇[1] 黄雯[1] 白静[1] 张梅[1] 

机构地区：[1]首都医科大学附属北京同仁医院肾内科、首都医科大学肾脏病学系,北京100730

出　　处：《医学研究杂志》2016年第4期85-89,共5页Journal of Medical Research

基　　金：首都医科大学基础-临床科研合作基金资助项目(11JL41)

摘　　要：目的探讨Chymase抑制对糖尿病大鼠心肌细胞凋亡的影响机制。方法 24只雄性SD大鼠随机分为正常对照组(NC组,n=7)、糖尿病组(DM组,n=7)、应用糜酶(Chymase)抑制剂的糖尿病组(DM+Chy-I组,n=10)。采用链脲佐菌素(streptozocin,STZ)单次腹腔注射方法建立糖尿病大鼠模型。其中DM+Chy-I组给予糜酶抑制剂[Suc-Val-Pro-Phe P-(OPh)2,1mg/(kg·d)],12周后处死,采用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测心肌细胞的凋亡指数(AI),Western blot法检测心肌caspase-3蛋白的表达,免疫组化检测心肌组织Chymase蛋白表达,采用黄嘌呤氧化酶法(羟胺法)测定心肌组织超氧化物歧化酶(SOD)活力,TBA(thiobarbituric acid)法测定丙二醛(MDA)含量。结果与NC组相比,DM组大鼠心肌细胞AI明显升高;caspase-3蛋白表达明显增加;MDA含量升高,SOD活力降低(P<0.05);与DM组相比,Chymase抑制剂使心肌细胞AI明显降低;caspase-3蛋白表达减少;MDA含量降低,SOD活力升高(P<0.05)。结论 Chymase抑制剂减少了糖尿病心肌病大鼠心肌细胞的凋亡可能是通过抑制caspase-3的表达和氧化应激损伤。Objective To investigate the mechanism of the effects on chymase inhibition in cardiomyocytes apoptosis. Methods Twenty-four male rats （Sprague-Dawley） were randomly divided into normal control group （NC group, n=7）, diabetes mellitus group （DM group, n=7） and Chy-I（chymase inhibitor） treatment group （DM＋Chy-I group, n=10）. Diabetes was induced in the rats （DM group, DM＋Chy-I group） by intraperitoneal injection of streptozocin （60mg/kg）.The rats in the DM＋Chy-I group were injected with chymase inhibitor Suc-Val-Pro-PheP-（OPh）2 1mg/（kg·d）, while the same volume of normal saline was administrated in NC group and DM group. Three groups of rats were sacrificed after 12 weeks of modeling, The apoptotic index（AI） of cardiomyocytes was detected by TUNEL. Expression of caspase-3 was measured by Western blot.The expression of chymase protein in the cardiomyocytes was detected by the method of immunochemistry. The content of maleic dialdehyde （MDA） in myocardium was assayed by thiobarbituric acid. The activities of superoxide dismutase（SOD） in myocardium was examined by xanthine oxidase （the hydroxylamine method）. Results The apoptosis index, caspase-3 proteins, content of maleic dialdehyde （MDA） were all increased and the activities of superoxide dismutase（SOD） was decreased in DM group. Chymase inhibition reversed the level of all in DM＋Chy-I group. Conclusion Chymase inhibition can reduce the myocardial cells apoptosis of diabetic rats. The mechanism maybe attenuates oxidative stress damage.

关 键 词：CHYMASE 糖尿病 细胞凋亡 氧化应激 

分 类 号：R587.1[医药卫生—内分泌]