糖尿病大鼠心肌缺血再灌注时Rev-erbα与NLRP3炎症小体的关系  

Relationship between Rev-erbαand NLRP3 inflammasome during myocardial ischemia-reperfusion in diabetic rats

作　　者：李璐[1] 邱珍[1] 田浩 夏中元[1] Li Lu;Qiu Zhen;Tian Hao;Xia Zhongyuan(Department of Anesthesiology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院麻醉科,武汉430060

出　　处：《中华麻醉学杂志》2022年第5期542-545,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81970722);中央高校基本科研业务费专项资金资助(2042021kf0113)。

摘　　要：目的评价糖尿病大鼠心肌缺血再灌注时Nr1D1核受体亚家族1,组D,成员1(Rev-erbα)与NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体的关系。方法SPF级健康成年雄性SD大鼠,体重210~240 g,采用腹腔注射1%链脲佐菌素60 mg/kg制备1型糖尿病模型。取非糖尿病大鼠18只,采用随机数字表法分为2组:非糖尿病假手术组(NS组,n=6)和非糖尿病心肌缺血再灌注组(NIR组,n=12)。取糖尿病大鼠30只,采用随机数字表法分为3组:糖尿病假手术组(DS组,n=6)、糖尿病心肌缺血再灌注组(DIR组,n=12)和糖尿病心肌缺血再灌注+Rev-erbα抑制剂SR8278组(DIR+SR组,n=12)。采用结扎左冠脉前降支30 min后再灌注120 min的方法建立心肌缺血再灌注损伤模型。DIR+SR组于缺血前1 h时经股静脉注射SR82782 mg/kg。再灌注结束时经采用ELISA法检测血清CK-MB、LDH和cTnI浓度,TTC法测心肌梗死面积百分比,HE染色法观察心肌组织病理学结果,Western blot法检测心肌组织Rev-erbα、NLRP3和IL-1β的表达水平。结果与假手术大鼠比较,心肌缺血再灌注大鼠血清CK-MB、LDH和cTnI浓度升高,心肌组织Rev-erbα、NLRP3和IL-1β表达上调(P<0.05),心肌组织病理学损伤明显;与NIR组比较,DIR组心肌梗死面积百分比、血清CK-MB、LDH和cTnI浓度升高,心肌组织Rev-erbα、NLRP3和IL-1β表达上调(P<0.05),心肌组织病理学损伤加重;与DIR组比较,DIR+SR组心肌梗死面积百分比、血清CK-MB、LDH和cTnI水平降低,心肌组织Rev-erbα、NLRP3和IL-1β表达下调(P<0.05),心肌组织病理学损伤减轻。结论Rev-erbα可促进NLRP3炎症小体活化,参与糖尿病大鼠心肌缺血再灌注损伤的病理生理机制。Objective To evaluate the relationship between nuclear receptor subfamily 1,group D,member 1(Rev-erbα)and NOD-like receptor protein 3(NLRP3)inflammasome during myocardial ischemia-reperfusion(I/R)in diabetic rats.Methods SPF-grade healthy male Sprague-Dawley rats,weighing 210-240 g,in which 1%streptozotocin 60 mg/kg was intraperitoneally injected to develop the model of type 1 diabetes mellitus.Eighteen non-diabetic rats were divided into 2 groups by the random number table method:non-diabetic sham operation group(NS group,n=6)and non-diabetic myocardial I/R group(NIR group,n=12).Thirty diabetic rats were divided into 3 groups by the random number table method:diabetic sham operation group(DS group,n=6),diabetic myocardial I/R group(DIR group,n=12),and diabetic myocardial I/R+Rev-erbαinhibitor SR8278 group(DIR+SR group,n=12).Myocardial I/R model was developed by ligation of left anterior descending coronary artery for 30 min followed by reperfusion for 120 min.In DIR+SR group,SR82782 mg/kg was injected via the femoral vein at 1 h before ischemia.At the end of reperfusion,blood samples from the right carotid artery were collected for determination of serum creatine kinase-MB(CK-MB),lactate dehydrogenase(LDH)and cardiac troponin I(cTnI)levels(by enzyme-linked immunosorbent assay).Then the rats were sacrificed,hearts were removed and myocardial tissues were obtained for determination of the percentage of myocardial infarct size(by TTC method)and expression of Rev-erbα,NLRP3 and IL-1β(by Western blot)and for microscopic examination of pathologic changes(by HE staining).Results Compared with sham-operated rats,the serum concentrations of CK-MB,LDH and cTnI were significantly increased,the expression of Rev-erbα,NLRP3 and IL-1βin myocardial tissues was up-regulated(P<0.05),and the pathological injury of myocardial tissues was obvious in myocardial I/R rats.Compared with NIR group,the percentage of myocardial infarct size and levels of serum CK-MB,LDH and cTnI were significantly increased,the expression of Rev-erb

关 键 词：糖尿病 心肌再灌注损伤 核受体亚家族1 D组 成员1 NLR家族 热蛋白结构域包含蛋白3 

分 类 号：R587.2[医药卫生—内分泌]