白藜芦醇调控细胞自噬通路改善糖尿病合并心肌缺血再灌注损伤的实验研究  

Experimental study of improvement effect of resveratrol on diabetes mellitus with ischemia reperfusion injury by regulating autophagy pathway

作　　者：杨礼[1] 徐麟皓[2] 陈肖[1] 屈献峰[1] 

机构地区：[1]台州市立医院麻醉科,浙江省台州318000 [2]杭州医学院解剖教研室

出　　处：《中国慢性病预防与控制》2018年第1期9-13,共5页Chinese Journal of Prevention and Control of Chronic Diseases

基　　金：国家自然科学基金项目(81601972);台州市科学技术局项目(162yw04)

摘　　要：目的研究白藜芦醇对糖尿病合并心肌缺血再灌注损伤大鼠自噬通路表达及心肌梗死面积的影响,进一步明确其药理机制。方法选取100只6~8周龄清洁级(SPF级)雄性SD大鼠为研究对象,随机分为4组,每组25只,分别为对照组、模型组、白藜芦醇组、三甲基腺嘌呤(3-MA)组,4组均采用高脂喂养结合腹腔注射链脲佐菌素法制备糖尿病模型,模型制备成功者给予药物干预。白藜芦醇组以10 mg/(kg·d)的白藜芦醇灌胃,3-MA组在白藜芦醇组的基础上尾静脉注射0.015 g/(kg·d)自噬抑制剂3-MA,对照组和模型组给予等量生理盐水刺激;7 d后,除对照组外,均采用冠状动脉结扎法制备心肌缺血再灌注模型。采用Med Lab生物信号采集处理系统测定药物干预前(基线值)、心肌缺血前、心肌缺血30 min和再灌注2 h的血流动力学数据,采用2,3,5-三苯基氯化四氮唑(TTC)染色实验测定心肌梗死面积,采用免疫蛋白印迹实验测定心肌组织自噬标志物蛋白表达。用SPSS 19.0统计软件进行t检验、方差分析和χ~2检验。结果各组大鼠的左心室收缩压(LVSP)、左心室等容收缩期(+)心室压力上升的最大速率(+dp/dt)及左心室等容舒张期(-)心室压力上升的最大速率(-dp/dt)基线值差异均无统计学意义(P>0.05);组别因素与时间因素对大鼠的LVSP、+dp/dt及-dp/dt水平均有影响(P<0.05),且组别与时间之间无交互作用(P>0.05)。组间比较结果显示,心肌缺血前,白藜芦醇组LVSP、+dp/dt及-dp/dt水平较其他各组有所升高;心肌缺血30 min和再灌注2 h,模型组和3-MA组LVSP、+dp/dt及-dp/dt水平低于对照组,白藜芦醇组LVSP、+dp/dt及-dp/dt水平高于模型组和3-MA组,差异均有统计学意义(P<0.05)。组内比较结果显示,心肌缺血前,白藜芦醇组LVSP、+dp/dt及-dp/dt水平高于基线值,差异均有统计学意义(P<0.05),其他各组与基线值比较,差异均无统计学意义(P>0.05);心肌缺血30 min和再灌注2 h时Objective To investigate the influence of resveratrol on autophagy pathway expression and myocardial infarction area in rats with diabetes mellitus and ischemia reperfusion injury, and to determine its pharmacological mechanism. Methods The subjects were 100 male SD rats（6-8 weeks old）, which were divided randomly into 4 groups（control, model, resveratrol and 3-MA groups）. The diabetes models were prepared by high fat diet and streptozotocin injection, and the successful models were given pharmacological intervention. The resveratrol group was given with 10 mg/（kg·d） resveratrol gavage, the 3-MA（autophagy inhibitor） group was given with 0.015 g/（kg·d） 3-MA by caudal vein injection on base of resveratrol. The control group and model group were given with equivalent normal saline stimulate. The model, resveratrol and 3-MA groups were prepared for ischemia-reperfusion models by ligating coronary artery. The haemodynamics data of all the rats before pharmacological intervention, before ischemia, 30 min after ischemia, and reperfusion for 2 h were detected by Med Lab bio-signals collecting-processing system. The myocardial infarction areas were measured by TTC staining experiment, the expression of autophagy marker proteins in cardiac muscle tissue was detected by western blot（WB） assay. The t test, χ~2 test and ANOVA were used to analyze the data, and the used software was SPSS 19.0. Results There were no significant differences in baseline values of LVSP, ＋dp/dt and-dp/dt among the four groups（P〉0.05）. There were the significant effects of group factor and time factor on the level of LVSP, ＋dp/dt and-dp/dt（P〈0.05）, but there was no interaction effect between group factor and time factor（P〉0.05）. Before ischemia, the LVSP, ＋dp/dt and-dp/dt levels of resveratrol group were significantly higher than those of other groups; in 30 min after ischemia and reperfusion for 2 h, the LVSP, ＋dp/dt and-dp/dt levels of model group and 3-MA group were significantly lower than tho

关 键 词：白藜芦醇 细胞自噬 糖尿病 心肌 缺血再灌注损伤 

分 类 号：R587.1[医药卫生—内分泌]