TGF-β1/Smad3信号通路调控MMP-13/TIMP-1蛋白表达在糖尿病膀胱纤维化中的作用研究  

The role of TGF-β1/Smad3 signaling pathway in regulating the expression of MMP-13/TIMP-1 protein in diabetic cystic fibrosis

作　　者：周海永 郁建迪[1] 闻华立 彭昆 颜俊锋 汪星 ZHOU Haiyong;YU Jiandi;WEN Huali;PENG Kun;YAN Junfeng;WANG Xing(Authors'address:Department of Urology,Zhejiang Hospital,Hangzhou 310013,China)

机构地区：[1]浙江医院泌尿外科,杭州310013

出　　处：《浙江医学》2023年第15期1598-1602,1620,共6页Zhejiang Medical Journal

基　　金：浙江省卫生健康科技计划项目(2022RC094)。

摘　　要：目的探讨转化生长因子β1(TGF-β1)/细胞信号转导分子3(Smad3)信号通路调控基质金属蛋白酶13(MMP-13)/基质金属蛋白酶抑制剂1(TIMP-1)蛋白表达在糖尿病膀胱纤维化中的作用。方法将80只雄性Wistar大鼠按随机数字表法分为糖尿病组和正常对照组,每组40只;另取20只糖尿病大鼠按随机数字表法分为TGF-β1敲除组和Smad3敲除组,每组10只。采用链脲佐素65 mg/kg一次性给药法建立糖尿病模型;采用Cas9 gene targeting技术分别敲除TGF-β1或Smad3基因。采用HE或VG染色法观察糖尿病组与正常对照组大鼠膀胱组织病理学形态,Western blot法检测并比较糖尿病组与正常对照组、糖尿病组与TGF-β1敲除组、糖尿病组与Smad3敲除组大鼠膀胱平滑肌组织中相关信号通路因子蛋白表达水平,包括TGF-β1、Smad3、MMP-13、TIMP-1、Ⅰ型胶原蛋白(colⅠ)、Ⅲ型胶原蛋白(colⅢ)等。结果HE及VG染色结果显示,糖尿病组大鼠膀胱组织胶原纤维明显增多。糖尿病组大鼠膀胱平滑肌组织中TGF-β1、Smad3、MMP-13、TIMP-1蛋白表达水平均明显高于正常对照组,MMP-13、TIMP-1、colⅠ、colⅢ蛋白表达水平均明显高于TGF-β1敲除组、Smad3敲除组,差异均有统计学意义(均P<0.05)。结论MMP-13、TIMP-1在糖尿病膀胱纤维化中呈高表达。TGF-β1/Smad3信号通路通过调控MMP-13/TIMP-1蛋白表达参与糖尿病膀胱纤维化的发生、发展。Objective To investigate the role of transforming growth factorβ1(TGF-β1)/signaling molecule 3(Smad3)signaling pathway in regulating the expression of matrix metalloproteinase 13(MMP-13)/tissue inhibitor of matrix metalloproteinase 1(TIMP-1)protein in diabetic cystic fibrosis.Methods Eighty male Wistar rats were randomly divided into control group and diabetic group,with 40 rats in each group.Another 20 diabetic rats were divided into TGF-β1 knockout group and Smad3 knockout group by random number table method,with 10 rats in each group.The diabetic model was induced by intraperitonian injection of streptozotocin 65 mg/kg.TGF-β1 and Smad3 genes were targeted by Cas9 gene targeting.HE or VG staining was used to observe the pathological morphology of the bladder tissue,and Western blot was used to detect the expression of TGF-β1/Smad3 signaling pathway related proteins,including TGF-β1,Smad3,MMP-13,TIMP-1,collagenⅠ(colⅠ)and collagenⅢ(colⅢ)in all groups.Results HE and VG staining results showed that the collagen fibers in bladder tissue of diabetic rats increased significantly.The protein expression levels of TGF-β1,Smad3,MMP-13 and TIMP-1 in bladder smooth muscle tissue of diabetic group were significantly higher than those of control group(all P<0.05),and the protein expression levels of MMP-13,TIMP-1,colⅠand colⅢwere significantly higher than those of TGF-β1 knockout group and Smad3 knockout group(all P<0.05).Conclusion MMP-13 and TIMP-1 are highly expressed in diabetic rats with bladder fibrosis.TGF-β1/Smad3 signaling pathway is involved in the occurrence and development of diabetic cystic fibrosis by regulating the expression of MMP-13/TIMP-1 protein.

关 键 词：糖尿病 膀胱 纤维化 基质金属蛋白酶13 基质金属蛋白酶抑制剂1 转化生长因子β1/细胞信号转导分子3信号通路 

分 类 号：R587.2[医药卫生—内分泌] R694[医药卫生—内科学]