心肌细胞外基质重塑对缝隙连接蛋白43及其Ser368位点磷酸化和电传导的影响  

Effects of myocardial extracellular matrix remodeling on connexin 43 and its Ser368 phosphorylation and electrical conduction

作　　者：宋雨婷 文春雷 李奕 柏雪 高鸿 胡廷菊 王子君 严旭 Song Yuting;Wen Chunlei;Li Yi;Bai Xue;Gao Hong;Hu Tingju;Wang Zijun;Yan Xu(School of Anesthesiology,Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Department of Anesthesiology,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China)

机构地区：[1]贵州医科大学麻醉学院,贵州省贵阳市550004 [2]贵州医科大学附属医院麻醉科,贵州省贵阳市550004

出　　处：《中国组织工程研究》2025年第29期6212-6218,共7页Chinese Journal of Tissue Engineering Research

基　　金：贵州省科技厅基础研究计划(自然科学类)项目(黔科合基础-ZK[2024]一般195),项目参与人:高鸿、文春雷、柏雪、宋雨婷。

摘　　要：背景:课题组前期研究发现,心肌低温缺血再灌注后缝隙连接蛋白43及其丝氨酸368位点磷酸化表达下降与心肌传导速度减慢和再灌注心律失常的发生密切相关。目的:观察低温缺血再灌注后心肌细胞外基质中膜型基质金属蛋白酶2、基质金属蛋白酶2和Ⅳ型胶原蛋白表达变化对心肌缝隙连接蛋白43及其丝氨酸368位点磷酸化表达和电传导的影响。方法:取16只SD大鼠心脏建立Langendorff体外心脏灌注模型,随机分为对照组(n=8)与低温缺血再灌注组(n=8),对照组用37℃Krebs-Henseleit液平衡灌注15 min后继续灌注37℃Krebs-Henseleit液90 min;低温缺血再灌注组用37℃Krebs-Henseleit液平衡灌注15 min,注射4℃Thomas液使心脏停搏60 min,心脏停搏期间周围用4℃Krebs-Henseleit液保护,停搏30 min时半量复灌4℃Thomas液,停搏结束后用37℃Krebs-Henseleit液再灌注30 min。于再灌注即刻至再灌注结束,记录心律失常发生情况、心脏复跳时间和心律失常持续时间;通过Mapping Lab多通道电生理标测系统采集平衡灌注15 min(T1)、再灌注15 min/继续灌注90 min(T2)、再灌注30 min/继续灌注105 min(T3)的传导速度、绝对不均匀性和不均匀性指数;再灌注结束后,采用免疫印迹法检测左心室肌膜型基质金属蛋白酶2、基质金属蛋白酶2、Ⅳ型胶原蛋白、缝隙连接蛋白43和丝氨酸368位点磷酸化缝隙连接蛋白43的蛋白表达。结果与结论:①对照组未发生心律失常;低温缺血再灌注组有6个心脏发生心律失常,心脏复跳时间与心律失常持续时间分别为(25.38±12.02),(158.67±67.68)s。②对照组T1、T2和T3时点的传导等时图均匀且方向规律,T2和T3时点的传导速度与T1时点比较无差异(P>0.05);低温缺血再灌注组T2和T3时点的传导等时图不均匀且方向不规律,传导速度慢于T1时点(P<0.01);低温缺血再灌注组T2和T3时点的传导速度慢于对照组(P<0.01);低温缺血再灌注组T2、T3时BACKGROUND:Our previous studies found that decreased expression of connexin 43 and its Ser368 phosphorylation after myocardial hypothermic ischemiareperfusion was closely associated with decreased cardiac conduction velocity and reperfusion arrhythmia.OBJECTIVE:To observe the effect of changes in membrane-type matrix metalloproteinase 2,matrix metalloproteinase 2 and collagen type Ⅳ on the expression of connexin 43 and its Ser368 phosphorylation and electrical conduction in the myocardial extracellular matrix after hypothermic ischemia-reperfusion.METHODS:Sixteen Langendorff extracorporeal cardiac perfusion models were successfully established from SD rats and randomly divided into a control group(n=8)and a hypothermic ischemia-reperfusion group(n=8).The control group was balanced perfused with 37℃ Krebs-Henseleit solution for 15 minutes and then continued to be perfused with 37℃ Krebs-Henseleit solution for 90 minutes.The hypothermic ischemia-reperfusion group was balanced perfused with 37℃ Krebs-Henseleit solution for 15 minutes,and then the heart was arrested for 60 minutes by injection of 4℃ Thomas solution.During the cardiac arrest,the periphery was protected by 4℃ Krebs-Henseleit solution.Half-volume 4℃ Thomas solution was reperfused 30 minutes after the arrest.After stopping the arrest,the heart was reperfused with 37℃ Krebs-Henseleit solution for 30 minutes.The occurrence of arrhythmias,rebeating time,and the duration of arrhythmias were recorded from the immediate time point to the end of the reperfusion period.Conduction velocity,absolute inhomogeneity,and inhomogeneity index were measured using the Mapping Lab multi-channel electrophysiological mapping system at the time of balanced perfusion for 15 minutes(T1),reperfusion for 15 minutes/continuous perfusion for 90 minutes(T2),and reperfusion for 30 minutes/continuous perfusion for 105 minutes(T3).The relative expression levels of membrane-type matrix metalloproteinase 2,matrix metalloproteinase 2,collagen type IV,connexin 43,and its Se

关 键 词：缝隙连接蛋白43 磷酸化缝隙连接蛋白43 再灌注心律失常 细胞外基质 膜型基质金属蛋白酶2 基质金属蛋白酶2 Ⅳ型胶原蛋白 工程化组织构建 

分 类 号：R459.9[医药卫生—治疗学] R363[医药卫生—临床医学] R541.7