摘要
目的:探究细胞焦亡在糖尿病大鼠心肌缺血再灌注损伤的变化及作用机制。方法:选择成年健康雄性SD大鼠,体重210-230g。大鼠糖尿病模型采用腹腔注射1%链脲佐菌素60mg/kg制备,取造模成功的SD大鼠40只,将其随机分为2组(n=20):糖尿病假手术组(DS组)、糖尿病心肌缺血再灌注组(DIR组);另取非糖尿病大鼠40只,将其随机分为2组(n=20):假手术组(NS组)、心肌缺血再灌注组(NIR组)。采用结扎左冠状动脉前降支30min再灌注120min的方法制备心肌缺血再灌注模型;NS组、DS组只穿线不结扎。再灌注结束后,采集动脉血样,检测肌酸激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)的活性;采用TTC法检测心肌梗死面积;HE染色观察心肌组织病理学变化;Western Blot法检测NLRP3、caspase-1、凋亡相关斑点样蛋白(ASC)和IL-1β在心肌组织的表达。结果:DS与NS相比,NLRP3、caspase-1、ASC和IL-1β蛋白表达增加;非糖尿病和糖尿病大鼠在心肌缺血再灌注时,CK-MB和LDH的活性增加,NLRP3、caspase-1、ASC和IL-1β蛋白表达增加;DIR组与NIR相比,心脏病理学损伤加重,心肌梗死面积增加,CK-MB和LDH活性增高,NLRP3、caspase-1、ASC和IL-1β蛋白表达增加。结论:糖尿病大鼠心肌对缺血再灌注敏感性增加,缺血再灌注损伤加重,其机制可能与NLRP3介导的caspase-1依赖性细胞焦亡的作用密切相关。
Objective：To evaluate the mechanism and effect of pyroptosis in myocardial ischemia-reperfusion in diabetic rats.Methods：Healthy adult male SD rats weighing 210-230 g were adopted in this study.Diabetes mellitus（DM）was induced by intraperitoneal injection of 1%streptozotocin60mg/kg and confirmed by blood glucose≥16.7mmol/L.Forty DM rats were randomly divided into 2groups（n=20each）：diabetes sham group（DS group）and diabetes myocardial ischemiareperfusion（I/R）group（DIR group）.Another forty healthy adult male SD rats were randomly divided into 2groups（n=20）：sham group（NS group）,myocardial I/R group（NIR group）.Myocardial I/R model was induced by 30 min occlusion of the left anterior descending branch of the coronary artery followed 120 min reperfusion.In group DS and group NS,left anterior de-scending was wearing without occluded.At the end of 120 min reperfusion,artery blood samples was collected for determination of serum creatine kinase（CK-MB）and lactate dehydrogenase（LDH）activity.TTC staining was used to assay myocardial infarction size.Pathology was observed under microscope.The expression of NLRP3,caspase-1,ASC and IL-1βin cardiac were measured by Western blot.Results：Compared with group NS,the expression of NLRP3,caspase-1,ASC and IL-1βprotein in group DS increased.The activities of CK-MB and LDH were significantly increased in diabetic and nondiabetic rats during myocardial I/R.The expressions of NLRP3,caspase-1,ASC and IL-1βprotein were also increased.Compared with group NIR,the infraction size,the activity of CK-MB and LDH,the expression of NLRP3,caspase-1,ASC and IL-1βprotein in group DIR were increased and with the pathological injury aggravated.Conclusion：The degree of myocardial ischemia-reperfusion injury is increasing in diabetic rats,the mechanism is associated with the NLRP3-induced caspase-1-independent pyroptosis to diabetes myocardial ischemia-reperfusion injury.
作者
邱珍
夏中元
周斌
吴洋
孟庆涛
冷燕
QIU Zhen XIA Zhongyuan ZHOU Bin WU Yang MENG Qingtao LENG Yan(Dept. of Anesthesiology, Renmin Hospital of Wuhan University, Wuhan 430060, Chin)
出处
《武汉大学学报（医学版）》
CAS
2017年第1期1-5,共5页
Medical Journal of Wuhan University
基金
国家自然科学基金资助项目(编号:81471844)
