RhoE基因表达改变在糖尿病心肌病心肌纤维化中的作用  

Role of RhoE gene expression changes in myocardial fibrosis in diabetic cardiomyopathy

作　　者：施凯佳 朱星霖 赵阳阳 柴金萱 申志华 郭峻莉 揭伟 Shi Kaijid;Zhu Xinglin;Zhao Yangyang;Chai Jinxuan;Shen Zhihud;Guo Junli;Jie Wei(School of Public Health,Key Laboratory of Tropical Translational Medicine of Ministry of Education,Hainan Medical University,Hainan Provincial Key Laboratory for Tropical Cardiovascular Diseases Research,Haikou 571199,China;Department of Pathophysiology,School of Basic Medicine Sciences,Guangdong Medical University,Zhanjiang 524023,China)

机构地区：[1]海南医科大学公共卫生学院,海南医科大学热带转化医学教育部重点实验室,海南省热带心血管病研究重点实验室,海口571199 [2]广东医科大学基础医学院病理生理学教研室,湛江524023

出　　处：《中华心血管病杂志》2025年第3期293-300,共8页Chinese Journal of Cardiology

基　　金：国家自然科学基金(82060053);海南省重点研发计划项目(ZDYF2020122,ZDYF2022SHFZ038)。

摘　　要：目的 探讨Ras同源基因家族成员E(RhoE)基因在糖尿病心肌病心肌纤维化中的作用及机制。方法 使用野生型SD大鼠腹腔注射链脲佐菌素溶液(STZ, 70 mg/kg)和等量柠檬酸钠溶液分别作为1型糖尿病(T1DM)组(n=15)和T1DM对照组(n=15)。以db/db自发性2型糖尿病(T2DM)小鼠和野生型C57BL/6J小鼠常规饲养8周分别作为T2DM组(n=5)和T2DM对照组(n=5)。使用RhoE基因全身敲除的杂合子SD大鼠腹腔注射STZ溶液(70 mg/kg)和等量柠檬酸钠溶液分别作为RhoE敲除T1DM组(n=5)和RhoE敲除对照组(n=5)。使用野生型SD大鼠尾静脉注射RhoE过表达的腺相关病毒9和腹腔注射STZ溶液(70 mg/kg)作为RhoE过表达T1DM组(n=5), 以尾静脉注射阴性对照病毒和腹腔注射等量柠檬酸钠溶液的野生型SD大鼠为RhoE过表达对照组(n=5)。成功建模后各组动物继续常规饲养6或8周即为实验终点。于实验终点行心脏超声检测各组动物心功能, 分析各组动物左心室射血分数(LVEF)和舒张早期二尖瓣血流速度峰值与晚期峰值比值(E/A比值)数据, 并采用Masson染色检测各组动物心肌组织中胶原纤维的沉积情况。采用Western blot法检测各组大鼠心肌组织中RhoE基因、Ⅰ型胶原、Ⅲ型胶原蛋白、Sma和Mad相关蛋白2/3(Smad2/3)和磷酸化Smad2/3蛋白的表达水平, 采用酶联免疫吸附试验法测定各组大鼠血清中转化生长因子-β1(TGF-β1)水平。结果 与各自对照组相比, T2DM组小鼠和T1DM组大鼠心脏组织中RhoE基因表达显著下调, 胶原纤维沉积更显著, LVEF和E/A比值均较低(P均<0.05)。与T1DM组比较, RhoE 敲除T1DM组大鼠心肌组织中Smad2/3磷酸化水平、Ⅰ型胶原和Ⅲ型胶原蛋白水平以及血清中TGF-β1水平均较高(P均<0.05), 胶原纤维沉积更明显(P<0.05)。此外, RhoE 过表达T1DM组大鼠的LVEF和E/A比值较T1DM组高(P均<0.05), 胶原纤维沉积较少(P<0.05)。结论 糖尿病通过抑制RhoE基因表达激活TGF-β1/Smads信号通路�Objective To explore the role and mechanism of Ras homolog gene family member E(RhoE)gene in myocardial fibrosis in diabetic cardiomyopathy.Methods Wild-type SD rats were intraperitoneally injected with streptozotocin solution(STZ,70 mg/kg)and an equal volume of sodium citrate solution to establish the type 1 diabetes mellitus(T1DM)group(n=15)and the T1DM control group(n=15),respectively.db/db spontaneous type 2 diabetes mellitus(T2DM)mice and wild-type C57BL/6J mice were conventionally housed for 8 weeks to establish the T2DM group(n=5)and the T2DM control group(n=5),respectively.Heterozygote SD rats with systemic knockout of the RhoE gene were intraperitoneally injected with STZ solution(70 mg/kg)and an equal volume of sodium citrate solution to establish the RhoE knockout T1DM group(n=5)and the RhoE knockout control group(n=5),respectively.Wild-type SD rats were injected with RhoE-overexpressing adeno-associated virus 9 through tail vein and intraperitoneally injected with STZ solution(70 mg/kg)to establish the RhoE overexpression T1DM group(n=5),while wild-type SD rats injected with negative control virus through tail vein and intraperitoneally injected with an equal volume of sodium citrate solution served as the RhoE overexpression control group(n=5).After successful modeling,all animals in each group were conventionally housed for an additional 6 or 8 weeks,which marked the experimental endpoint.At the experimental endpoint,echocardiography was performed to assess cardiac function of animals in each group,and left ventricular ejection fraction(LVEF)and the ratio of early to late diastolic transmitral flow velocity(E/A ratio)were analysed.Masson staining was used to detect collagen fiber deposition in myocardial tissue of animals in each group.Western blot analysis was conducted to detect the expression levels of RhoE gene,type I collagen,type IIl collagen,Smad2/3,and phosphorylated Smad2/3 protein in myocardial tissue of rats.Enzyme-linked immunosorbent assay was used to measure the levels of transforming

关 键 词：糖尿病 糖尿病心肌病 Ras同源基因家族成员E 心肌纤维化 转化生长因子β1/Smads信号 

分 类 号：R58[医药卫生—内分泌]