Cx43对胆总管结扎大鼠心肌损伤的负向调控作用及其机制  

The negative regulatory effect of connexin 43 on myocardial injury in rats with common bile duct ligation and its mechanism

作　　者：王晓玉 吕琳[1] 杨瀚 祝琳[1] 董胜男 董河[1] WANG Xiaoyu;LYU Lin;YANG Han;ZHU Lin;DONG Shengnan;DONG He(Department of Anesthesiology,The Affiliated Hospital of Qingdao University,Qingdao 266075,China)

机构地区：[1]青岛大学附属医院麻醉科,山东青岛266075 [2]青岛大学医学部 [3]青岛大学附属医院泌尿外科

出　　处：《精准医学杂志》2024年第4期304-308,共5页Journal of Precision Medicine

基　　金：国家自然科学基金项目(81902001)。

摘　　要：目的探讨缝隙连接蛋白43(Cx43)在胆总管结扎(CBDL)引起的大鼠心肌损伤中的作用及其分子机制。方法将18只SD大鼠随机分为假手术组、CBDL组及Cx43抑制剂组,每组6只。假手术组大鼠行胆总管游离术,CBDL组大鼠游离、结扎并切断胆总管,Cx43抑制剂组大鼠术前24 h通过尾静脉单次注射25 mg/kg的Gap 26后,游离、结扎并切断胆总管。三组大鼠饲养至术后第14天时采用心脏超声检查评估心脏射血分数(EF)、缩短分数(FS)和心率(HR);随后测定三组大鼠血清总胆红素(TBil)、苯丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、乳酸脱氢酶(LDH)、肌酸激酶-MB(CK-MB)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)活性及丙二醛(MDA)水平;取三组大鼠心脏组织行HE染色和Masson染色;采用免疫印迹实验检测心脏组织中Cx43、Wnt 3α、β-catenin蛋白的水平。结果心脏超声检查结果显示,CBDL组、Cx43抑制剂组大鼠心脏EF、FS、HR均显著低于假手术组(F=22.95~43.55,t=5.10~9.32,P<0.05)。血生化及氧化应激指标检测结果显示,与假手术组相比,CBDL组、Cx43抑制剂组大鼠血清中TBil、AST、ALT、LDH、CK-MB、MDA、GSH-PX水平显著升高,SOD的水平显著降低(F=19.29~100.60,t=3.67~13.72,P<0.05)。HE染色及Masson染色结果显示,CBDL组和Cx43抑制剂组大鼠发生心肌损伤及心肌间质纤维化。免疫印迹实验结果显示,与假手术组相比,CBDL组和Cx43抑制剂组大鼠心脏组织中Cx43水平显著下降,Wnt 3α、β-catenin水平显著升高(F=28.50~70.07,t=4.32~11.79,P<0.05);与CBDL组相比,Cx43抑制剂组大鼠心脏组织中Cx43水平显著下降,Wnt 3α、β-catenin水平显著升高(t=4.95~5.20,P<0.05)。结论CBDL大鼠心脏组织中Cx43下调可负向调控激活Wnt 3α/β-catenin通路,导致大鼠心肌组织损伤加剧及心脏功能下降。Objective To investigate the role and molecular mechanism of connexin 43(Cx43)in rats with myocardial injury induced by common bile duct ligation(CBDL).Methods A total of 18 Sprague-Dawley rats were randomly divided into sham-operation group,CBDL group,and Cx43 inhibitor group,with 6 rats in each group.The rats in the sham-operation group were given isolation of the common bile duct,those in the CBDL group were given isolation,ligation,and cutting of the common bile duct,and those in the Cx43 inhibitor group were given a single injection of 25 mg/kg Gap 26 via the caudal vein at 24 h before surgery,followed by the isolation,ligation,and cutting of the common bile duct.The rats in all three groups were reared to day 14 after surgery,and echocardiography was used to evaluate ejection fraction(EF),fractional shortening(FS),and heart rate(HR);the three groups were measured in terms of serum total bilirubin(TBil),alanine aminotransferase(ALT),aspartate aminotransferase(AST),lactate dehydrogenase(LDH),creatine kinase-MB(CK-MB),superoxide dismutase(SOD),glutathione peroxidase(GSH-PX)activity,and malondialdehyde(MDA);HE staining and Masson staining were performed for heart tissues samples collected from the three groups;Western blotting was used to measure the levels of Cx43,Wnt 3α,andβ-catenin in heart tissue.Results Echocardiography showed that the CBDL group and the Cx43 inhibitor group had significantly lower cardiac EF,FS,and HR than the sham-operation group(F=22.95-43.55,t=5.10-9.32,P<0.05).Measurement of blood biochemical parameters and oxidative stress indicators showed that compared with the sham-operation group,the CBDL group and the Cx43 inhibitor group had significant increases in serum TBil,AST,ALT,LDH,CK-MB,MDA,and GSH-PX and a significant reduction in SOD(F=19.29-100.60,t=3.67-13.72,P<0.05).HE staining and Masson staining showed myocardial injury and myocardial interstitial fibrosis in the CBDL group and the Cx43 inhibitor group.Western blotting showed that compared with the sham-operation group,the CBDL

关 键 词：心脏损伤 胆道疾病 胆总管 结扎术 纤维化 连接蛋白43 Wnt信号通路 Β连环素 

分 类 号：R541[医药卫生—心血管疾病] R575[医药卫生—内科学]