咪达唑仑对2型糖尿病心肌缺血再灌注大鼠心肌损伤、氧化应激和炎症反应的影响  

Effects of Midazolam on Myocardial Injury,Oxidative Stress and Inflammatory Response in Type 2 Diabetic Myocardial Ischemia-reperfusion Rats

作　　者：李前辉[1] 郭浩翔 谢小娟[1] 张宜林[1] LI Qianhui;GUO Haoxiang;XIE Xiaojuan;ZHANG Yilin(The First Affiliated Hospital of Henan University of Science & Technology,Luoyang 471023,Henan,China)

机构地区：[1]河南科技大学第一附属医院,河南洛阳471023

出　　处：《中西医结合心脑血管病杂志》2021年第16期2735-2740,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：河南省教育厅自然科学研究计划项目(No.2015B20235)。

摘　　要：目的观察咪达唑仑对2型糖尿病心肌缺血再灌注大鼠心肌损伤、炎症反应和氧化应激的影响。方法将Wistar雄性大鼠随机分为对照组和糖尿病组。观察组通过高脂饮食和链脲霉素方法诱导大鼠2型糖尿病模型。将糖尿病大鼠随机分为糖尿病假手术组(DS组)、糖尿病心肌缺血再灌注组(DI/R组)、糖尿病心肌缺血再灌注大鼠给药组(DI/RM组);将对照组大鼠随机分为假手术组(sham组)、心肌缺血再灌注组(I/R组)、心肌缺血再灌注给药组(I/RM组)。结扎冠状动脉获得局部缺血构建心肌缺血再灌注模型。缺血给药前20 min,I/RM组和DI/RM组大鼠静脉注射咪达唑仑0.75 mg/kg,并以1 mg/(kg·h)微注射泵维持至再灌注结束。FDP-1 HRV和BRS分析系统测定心功能;酶联免疫吸附测定法(ELISA)检测血清肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)、心肌肌钙蛋白I(cTnI)、肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-6、IL-1β和心肌组织丙二醛(MDA)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)表达水平;苏木精-伊红(HE)染色观察心肌损伤;流式细胞仪检测细胞凋亡;蛋白印迹法检测核因子E2相关因子2(Nrf2)、还原型烟酰胺腺嘌呤二核苷酸磷酸醌氧化还原酶-1(NQO-1)、血红素氧合酶-1(HO-1)蛋白表达。结果与DI/R组比较,DI/RM组大鼠心脏左室收缩压(LVSP)、心率(HR)和冠状动脉流量(CF)升高(P<0.01),左室舒张末期压力(LVEDP)及血清LDH和CK-MB活性、cTnI含量降低(P<0.01),心肌细胞逐渐恢复整齐排列,细胞变性坏死减少,心肌细胞凋亡率降低(P<0.01),GSH含量和SOD活性升高(P<0.01),MDA含量降低(P<0.01),血清TNF-α、IL-6、IL-1β含量减少(P<0.01),心肌细胞Nrf2、NQO-1和HO-1表达上调(P<0.01)。结论咪达唑仑可减轻2型糖尿病心肌缺血再灌注大鼠心肌损伤,抑制氧化应激和炎症反应,可能与调节Nrf2通路有关。Objective To observe the effects of midazolam on myocardial injury,inflammation and oxidative stress myocardial ischemia-reperfusion in type 2 diabetic rats.Methods The rats were randomly divided into the control group and the diabetic group.The rat model of type 2 diabetes was induced by high-fat diet and streptozotocin.Diabetic rats were randomly divided into diabetic sham operation group(DS group),diabetic myocardial ischemia-reperfusion group(DI/R group),and diabetic myocardial ischemia-reperfusion medication group(DI/RM group).Healthy male Wistar rats were randomly divided into sham operation group(sham group),myocardial ischemia-reperfusion group(I/R group)and myocardial ischemia-reperfusion medication group(I/RM group).The model of myocardial ischemia-reperfusion model was constructed by ligation of coronary artery to obtain ischemia.20 minutes before administration of ischemia,the rats in the I/RM group and DI/RM group were intravenously injected with midazolam 0.75 mg/kg,and were maintained at 1 mg/(kg·h)microinjection pump until reperfusion.Cardiac function was tested by FDP-1 HRV and BRS analysis system.Serum creatine kinase isoenzyme(CK-MB),lactate dehydrogenase(LDH),cardiac troponin I(cTnI),tumor necrosis factorα(TNF-α),interleukin(IL)-6,IL-1β,and malondialdehyde(MDA),glutathione(GSH),and superoxide dismutase(SOD)were detected by enzyme-linked immunosorption(ELISA).The myocardial injury was observed by hematoxylin-eosin(HE)staining.Apoptosis was detected by flow cytometry.The expressions of nuclear factor E2 related factor 2(Nrf2),reduced nicotinamide adenine dinucleotide phosphate quinone oxidoreductase 1(NQO-1)and heme oxygenase 1(HO-1)proteins were detected by Western Blotting.Results Compared with the DI/R group,left ventricular systolic pressure(LVSP),heart rate(HR),and coronary artery flow(CF)in the DI/RM group were increased(P<0.01),while left ventricular end diastolic pressure(LVEDP),serum LDH and CK-MB activities and cTnI content were decreased(P<0.01).Cardiomyocytes gradually recovered

关 键 词：心肌缺血再灌注 2型糖尿病 心肌损伤 咪达唑仑 炎症反应 氧化应激 大鼠 实验研究 

分 类 号：R54[医药卫生—心血管疾病]