左归降糖舒心方调控自噬对糖尿病转基因MKR鼠心肌损伤的影响  

Effects of Zuogui Jiangtang Shuxin Recipe on Autophagy on Myocardial Injury in Diabetes Transgenic MKR Mice

作　　者：田娜 喻嵘[2] 张薇薇[3] 刘秀[1] 肖瑶[3] 陈大光 TIAN Na;YU Rong;ZHANG Weiwei;LIU Xiu;XIAO Yao;CHEN Daguang(Hunan University of Chinese Medicine,Changsha 410000,China;The First Hospital of Hunan University of Chinese Medicine,Changsha 410000,China;Hunan Provincial People s Hospital,Changsha 410000,China;People s Hospital of Ningxiang,Hunang Ningxiang 410600,China)

机构地区：[1]湖南中医药大学,长沙410000 [2]湖南中医药大学第一附属医院,长沙410000 [3]湖南省人民医院,长沙410000 [4]宁乡市人民医院,湖南宁乡410600

出　　处：《中国中医基础医学杂志》2023年第5期747-752,共6页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金面上项目(82074400);湖南省科学厅重点研发计划项目(2020SK2101);湖南省教育厅科技计划项目创新平台开放基金项目(20K094)。

摘　　要：目的 通过观察左归降糖舒心方对骨骼肌胰岛素样生长因子1及胰岛素双受体功能缺失(mouse overexpressing a dominant-negative IGF-I receptor specifically in muscle,MKR)糖尿病心肌病小鼠的自噬标志蛋白Beclin-1表达的影响,研究调控自噬水平对糖尿病心肌损伤的保护作用机制。方法 C57 BL小鼠设作正常组,普通饲料喂养,MKR小鼠采用高脂饲养(high-fat diet,HFD)联合多次链脲佐菌素(streptozocin,STZ)腹腔注射造模后,随机分为模型组、左归降糖舒心方组(中药组)、雷帕霉素组、3-甲基腺嘌呤(3-Methyladenine,3-MA)组、盐酸二甲双胍片+马来酸依那普利片组(西药组),药物干预28 d。记录小鼠一般情况及空腹血糖(fasting blood glucose,FBG);ELISA检测肌钙蛋白I(cardiac troponin I,cTnI)和肌酸激酶同工酶(creatine kinase isoenzymes,CKMB)以评价心肌损伤程度;透射电镜观察心肌形态结构的改变及自噬体;免疫组化检测心肌胶原蛋白Ⅰ、胶原蛋白Ⅲ以评估心肌纤维化程度;免疫荧光检测自噬标志蛋白Beclin-1表达。结果 与正常组比较,模型组小鼠多饮多食多尿,FBG升高明显(P<0.01),cTnI、CK-MB水平及胶原蛋白Ⅰ、Ⅲ的阳性表达均显著升高(P<0.01),Beclin-1蛋白表达明显下降(P<0.01);电镜下心肌细胞紊乱,各细胞器形态显著改变。与模型组比较,中药组小鼠FBG降低(P<0.01),cTnI、CK-MB水平与胶原蛋白Ⅰ、胶原蛋白Ⅲ表达均下降(P<0.01),Beclin-1蛋白表达增强(P<0.01);心肌细胞形态明显改善。中药组小鼠胶原蛋白Ⅰ、Ⅲ表达显著低于西药组(P<0.01)。与模型组比较,3-MA组小鼠Beclin-1蛋白表达减弱(P<0.01), cTnI、CK-MB水平及胶原蛋白Ⅰ、Ⅲ表达水平升高(P<0.01)。与模型组比较,雷帕霉素组小鼠Beclin-1蛋白表达增强(P<0.01), cTn I、CK-MB水平及胶原蛋白Ⅰ、Ⅲ表达水平下降(P<0.01)。结论 左归降糖舒心方可能通过上调自噬水平,减轻MKR糖尿病心肌病小鼠的心肌纤维化、�Objective By examining the expression of autophagy marker protein Beclin-1 in the MKR diabetic cardiomyopathy model mouse,to investigate the protective mechanism of Zuogui Jiangtang Shuxin recipe in protecting diabetic myocardial injury.Methods C57BL mice were set up as the normal group,fed with ordinary feed.MKR mice were modeled with high-fat feeding and intraperitoneal injections of streptozocin(STZ)repeatedly.Then the modeled mice were randomly divided into model group,Zuogui Jiangtang Shuxin recipe group(traditional Chinese medicine group),Rapamycin Group,3-methyladenine(3-MA)group,and Metformin Hydrochloride Tablets combined Enalapril Maleate Tablets group(western medicine group).Each group was given the corresponding intervention for 28 days.Observed the general condition of mice,and recorded fasting blood glucose(FBG)of all groups during the experiment.ELISA was used to test Cardiac troponin I(cTnI)and Creatine Kinase Isoenzyme(CK-MB)to assess myocardial damage of mice,transmission electron microscope was used to observe the changes of myocardial morphological structure and autophagosomes,immunohistochemistry was used to detect myocardial collagenⅠand collagenⅢto assess the degree of myocardial fibrosis,immunofluorescence was used to observe the expression of autophagy marker Beclin-1 protein.Results Compared with the normal group,in the model group,the mice developed polydipsia,polyphagia and polyuria,and FBG of mice increased significantly(P<0.01),the levels of cTnI,CK-MB,collagenⅠandⅢwere obviously increased(P<0.01),and the expressions of Beclin-1 protein were obviously decreased(P<0.01).Under the electron microscope,the cardiomyocytes were disordered,and the morphology of the organelles damaged significantly.Compared with the model group,the FBG of mice in the traditional Chinese medicine group was decreased(P<0.01),the levels of cTnI and CK-MB were decreased(P<0.01),the expressions of collagen I and collagenⅢwere reduced(P<0.01),and the expression of Beclin-1 protein was enhanced(P<0.01),ca

关 键 词：糖尿病心肌病 左归降糖舒心方 自噬 BECLIN-1 MKR小鼠 

分 类 号：R285.5[医药卫生—中药学]