远隔缺血预处理对大鼠再灌注性心律失常的影响及Shh/Gli1信号通路在其中的作用  

Effect of remote ischemic preconditioning on reperfusion arrhythmia in rats and the role of Shh/Gli1 signaling pathway

作　　者：郭高锋 容雄飞 徐红党 张加强 Guo Gaofeng;Rong Xiongfei;Xu Hongdang;Zhang Jiaqiang(Department of Anesthesiology and Perioperative Medicine,Henan Provincial People's Hospital,Zhengzhou University People's Hospital,Henan University People's Hospital,Zhengzhou 450003,China)

机构地区：[1]河南省人民医院麻醉与围术期医学科,郑州大学人民医院,河南大学人民医院,450003

出　　处：《中华麻醉学杂志》2019年第11期1306-1310,共5页Chinese Journal of Anesthesiology

摘　　要：目的评价远隔缺血预处理对大鼠再灌注性心律失常的影响及音猬因子(Shh)/胶质瘤相关癌基因同源物1(Gli1)信号通路在其中的作用。方法清洁级健康雄性SD大鼠40只,6~8周龄,体重220~280 g,采用随机数字表法分为4组(n=10):假手术组(Sham组)、心肌缺血再灌注组(I/R组)、远隔缺血预处理组(RIPC组)和环巴胺+远隔缺血预处理组(CYC+RIPC组)。采用结扎左冠状动脉左前降支30 min再灌注2 h的方法建立心肌缺血再灌注损伤模型。RIPC组于右侧腹股沟处以止血带阻断下肢血流10 min后,松开止血带恢复灌注30 min,随后制备心肌缺血再灌注损伤模型。CYC+RIPC组于心肌缺血前30 min腹腔注射Shh特异性抑制剂环巴胺10 mg/kg,随即进行远隔缺血预处理。记录室性心律失常发生情况。再灌注120 min时取血样,采用7600型全自动生化分析仪测定血清CK-MB和cTnI浓度。取血结束后,取缺血区心肌组织,采用ELISA法检测TNF-α含量,观察心肌组织病理学形态,采用TUNEL法确定心肌细胞凋亡指数(AI),采用免疫荧光法观察心肌组织Shh和Gli1的表达部位,采用Western blot法测定心肌组织Shh和Gli1的表达水平。结果与Sham组比较,I/R组心律失常发生率和发作次数升高,发作总时间延长,心律失常评分升高,血清cTnI和CK-MB浓度、心肌TNF-α含量及AI升高,Shh和Gli1表达上调(P<0.05),心肌组织病理学损伤严重;与I/R组比较,RIPC组心律失常发生率和发作次数降低,发作总时间缩短,血清cTnI和CK-MB浓度、心肌TNF-α含量及AI降低,Shh和Gli1表达上调(P<0.05),心肌组织病理学损伤减轻,CYC+RIPC组上述指标差异无统计学意义(P>0.05);与RIPC组比较,CYC+RIPC组心律失常发生率和发作次数升高,发作总时间延长,血清cTnI和CK-MB浓度、心肌TNF-α含量及AI升高,Shh和Gli1表达下调(P<0.05)。结论远隔缺血预处理可减轻大鼠再灌注性心律失常,机制与激活Shh/Gli1信号通路有关。Objective To evaluate the effect of remote ischemic preconditioning(RIPC)on reperfusion arrhythmia and the role of Sonic hedgehog(Shh)/glioma associated oncogene homolog 1(Gli-1)signaling pathway in rats.Methods Forty clean-grade healthy male SD rats,aged 6-8 weeks,weighing 220-280 g,were divided into 4 groups(n=10 each)using a random number table method:sham operation group(Sham group),myocardial I/R group(I/R group),remote ischemic preconditioning group(RIPC group)and cyclopamine+remote ischemic preconditioning group(CYC+RIPC group).Myocardial I/R was produced by occlusion of left anterior descending branch of coronary artery for 30 min followed by 2 h reperfusion in anesthetized rats.In RIPC group,an elastic rubber tourniquet was used to block the lower limb blood flow for 10 min in the right groin and then released for 30-min reperfusion,and then the myocardial I/R model was established.In CYC+RIPC group,a Shh specific inhibitor cyclopamine 10 mg/kg was intraperitoneally injected at 30 min before ischemia,and then RIPC was performed.The development of ventricular arrhythmia was recorded.Blood samples were taken at 120 min of reperfusion for determination of the concentrations of serum creatine kinase-MB(CK-MB)and cardiac troponin I(cTnI)using a 7600 automatic biochemical analyzer.The myocardial tissues in the ischemic area were taken after collecting blood for examination of pathological changes and for determination of tumor necrosis factorα(TNF-α)content(by enzyme-linked immunosorbent assay),cell apoptosis(using TUNEL),expression sites of Shh and Gli1(by immunofluorescence)and expression of Shh and Gli1(by Western blot).AI was calculated.Results Compared with Sham group,the incidence and times of arrhythmia were significantly increased,the total duration of arrhythmia was prolonged,the arrhythmia score was increased,the serum cTnI and CK-MB concentrations,myocardial TNF-αcontent and AI were increased,the expression of Shh and Gli1 was up-regulated(P<0.05),and the pathological changes of myocardial tissue

关 键 词：缺血预处理 四肢 猬蛋白质类 心肌再灌注损伤 心律失常 心性 

分 类 号：R614[医药卫生—麻醉学]