Gja1基因重组慢病毒对糖尿病豚鼠膀胱病变模型中缝隙连接蛋白43蛋白及mRNA表达的影响  

Effect of Gja1 gene recombinant lentivirus on Cx43 protein and mRNA expression in a diabetic guinea pig bladder model

作　　者：张永强 潘凤 孙鹏 谭明辉 轩留明 王勤章[1] Zhang Yongqiang;Pan Feng;Sun Peng;Tan Minghui;Xuan Liuming;Wang Qinzhang(Department of Urology,Shihezi University School of Medicine,Shihezi 832000,Xinjiang Uygur Autonomous Region,China;Department of Pathology,The First Affiliated Hospital,Shihezi University School of Medicine,Shihezi 832000,Xinjiang Uygur Autonomous Region,China;Department of Pathology,Shihezi University School of Medicine,Shihezi 832000,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]石河子大学医学院第一附属医院泌尿外科,新疆维吾尔自治区石河子市832000 [2]石河子大学医学院第一附属医院病理科,新疆维吾尔自治区石河子市832000 [3]石河子大学医学院病理系,新疆维吾尔自治区石河子市832000

出　　处：《中国组织工程研究》2023年第8期1161-1165,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(8196030314),项目负责人:王勤章。

摘　　要：背景:缝隙连接蛋白43又被称为Gja1蛋白,Gja1基因重组慢病毒对糖尿病膀胱引起的收缩功能障碍可能有改善作用。目的:构建豚鼠糖尿病膀胱病变模型,研究经尿道灌注Gja1基因重组慢病毒药物的方式,观察对受损膀胱逼尿肌细胞表面缝隙连接蛋白43蛋白和mRNA表达的影响。方法:选取80只鼠龄及体质量相近的健康豚鼠,随机原则挑选15只豚鼠常规饲养,设为正常组;剩余65只高糖高脂饮食喂养4周后给予腹腔注射1%链脲佐菌素200 mg/kg建立糖尿病豚鼠膀胱模型;尿动力学筛选出符合糖尿病膀胱病变豚鼠模型,依据随机分组原则分为3组:空白组(n=12)、对照组(n=12)、实验组(n=12)。空白组经尿道灌注0.2 mL磷酸缓冲盐溶液;对照组经尿道灌注0.2 mL空载基因重组慢病毒;实验组经尿道灌注0.2 mL Gja1基因重组慢病毒。每组在转染后第2,7,14,28天分别处死3只豚鼠,通过免疫组织化学染色法观察豚鼠膀胱组织缝隙连接蛋白43蛋白的表达及分布,分别采用Western-Blot和q RT-PCR检测缝隙连接蛋白43蛋白和mRNA的表达水平。结果与结论:①经尿道灌注Gja1基因重组慢病毒后,实验组较空白组及对照组的缝隙连接蛋白43蛋白和mRNA表达水平均明显升高(P<0.01,P<0.01),且基因重组慢病毒在转染后第14天缝隙连接蛋白43蛋白和mRNA表达最佳;空白组较对照组缝隙连接蛋白43蛋白和mRNA表达水平差异无显著性意义(P>0.01);②结论:经尿道灌注Gja1基因可以稳定表达于膀胱组织中,并能上调缝隙连接蛋白43蛋白和mRNA的表达,有利于修复细胞间受损的信号转导及物质交换通路。BACKGROUND:Connexin 43 is also known as Gja1 protein.Gja1 recombinant lentivirus may improve bladder contractile dysfunction in diabetes.OBJECTIVE:To establish a guinea pig model of diabetic cystopathy,study the way of transurethral infusion of Gja1 recombinant lentiviral drugs,and observe the effect on the expression of connexin 43 protein and mRNA on the surface of detrusor muscle cells in the damaged bladder.METHODS:Eighty healthy guinea pigs of similar age and body mass were selected,15 of which were randomly selected and conventionally fed and the remaining 65 were given intraperitoneal injection of 1%streptozotocin(200 mg/kg)to establish a guinea pig model of diabetic cystopathy.Urodynamics screening was performed to identify guinea pigs with diabetic cystopathy,which were further randomized into three groups:blank group(n=12),control group(n=12)and experimental group(n=12).The blank group was perfused with 0.2 mL of phosphate buffered saline,the control group was perfused with 0.2 mL of empty recombinant lentivirus,and the experimental group was perfused with 0.2 mL of Gja1 recombinant lentivirus.Three guinea pigs from each group were killed at each observation time point(2,7,14,and 28 days)after transfection.Expression and distribution of connexin 43 protein in the bladder were observed by immunohistochemical staining,and the protein and mRNA levels of connexin 43 were detected by western blot and qRT-PCR assays,respectively,RESULTS AND CONCLUSION:After transurethral infusion of Gja1 recombinant lentivirus,the expression levels of connexin 43 protein and mRNA were significantly higher in the experimental group than the blank and control groups(P<0.01,P<0.01),and the best expression levels of connexin 43 protein and mRNA were found at 14 days after transfection with Gja1 recombinant lentivirus.There were no significant differences in the expression levels of connexin 43protein and mRNA between the blank and control groups(P>0.01).To conclude,Gja1 gene via transurethral perfusion can stably express in bladd

关 键 词：Gja1基因 重组慢病毒 糖尿病膀胱 CX43蛋白 

分 类 号：R694[医药卫生—泌尿科学] R446[医药卫生—外科学] R318[医药卫生—临床医学]