RhoA/ROCK通路在2型糖尿病大鼠心肌纤维化形成中的作用及法舒地尔的干预效果  

Roles of RhoA/ROCK pathway in myocardial fibrosis formation in rats with type 2 diabetes mellitus and intervention effect of fasudil

作　　者：黄柳[1] 贾妍[1] 郭炳彦[1] 刘素云[1] 李拥军 HUANG Liu;JIA Yan;GUO Binyan;LIU Suyun;LI Yongjun(Department of the Fourth Internal Medicine-Cardiovascular,The Second Hospital of Hebei Medical University,Shjiazhuang 050000,China)

机构地区：[1]河北医科大学第二医院心血管内四科,石家庄050000

出　　处：《新疆医科大学学报》2020年第6期701-706,共6页Journal of Xinjiang Medical University

基　　金：国家自然科学基金青年科学基金(81400217);国家自然科学基金面上项目(81570345);河北省医学科学研究重点课题计划(20180329)。

摘　　要：目的研究RhoA/ROCK通路在2型糖尿病(T2MD)大鼠心肌纤维化形成中的作用及法舒地尔的干预效果。方法将30只SD大鼠随机分为正常组、模型组、法舒地尔干预组,每组10只,除正常组外,其他组均给予高糖高脂饲料,并腹腔注射链脲佐菌素(STZ)制备糖尿病模型,造模完成24 h后,法舒地尔干预组给予盐酸法舒地尔注射液10 mg/kg注射,1次/d,模型组和正常组大鼠给予等量生理盐水注射,持续4周,4周后处死大鼠,取心脏进行称量并计算心脏肥厚指数(HWI),用10%甲醛固定后,光镜及电镜下观察心肌纤维化情况;检测心肌中胶原含量及胶原面积;利用qRT-PCR检测心肌组织中I型前胶原、Rho激酶1(ROCK1)、转化生长因子-β1(TGF-β1)mRNA的相对表达水平;利用Western blot检测心肌磷酸化肌球蛋白磷酸酯酶靶点亚单位1(p-MYPT1)、细胞自噬相关蛋白(LC3-I、LC3-II、p62)表达情况。结果与正常组相比,模型组大鼠光、电镜可见心肌组织病理纤维化严重,同时HWI、胶原含量、胶原面积均升高,I型前胶原、ROCK1、TGF-β1的mRNA相对表达水平升高,LC3-II/LC3-I比例降低,p62蛋白表达水平升高,差异均有统计学意义(P<0.05);与模型组相比,法舒地尔干预组大鼠光、电镜下心肌组织病理纤维化减轻,同时HWI、胶原含量、胶原面积均降低,I型前胶原、ROCK1、TGF-β1的mRNA相对表达水平降低,LC3-II/LC3-I比例升高,p62蛋白表达水平降低,差异均有统计学意义(P<0.05)。结论T2MD通过激活RhoA/ROCK通路上调心肌细胞中促纤维化细胞因子表达及抑制心肌细胞自噬促进心肌纤维化,法舒地尔可以通过抑制RhoA/ROCK通路干预T2MD引起的心肌纤维化。Objective To study molecular mechanism of myocardial fibrosis in type 2 diabetes mellitus(T2DM)rats and intervention effect of fasudil.Methods A total of 30 SD rats were randomly divided into normal group,model group and fasudil intervention group with 10 cases in each group.Except the normal group,the other groups were given high glucose and high fat diet and intraperitoneal injection of streptozotocin(STZ)to prepare diabetes mellitus models.At 24h after modeling,fasudil intervention group was given 10 mg/kg fasudil hydrochloride injection,once/d,while the model group and normal group were given equivalent normal saline for 4 weeks.The heart weight and heart weight index(HWI)were recorded and calculated.Myocardial fibrosis was observed under light and electron microscope.The content and area of collagen in myocardial homogenate were detected.The relative expression levels of type I procollagen,Rho kinase(ROCK1)and transforming growth factor-β1(TGF-β1)mRNA in myocardial tissue were detected by qRT-PCR.The expression of phosphorylated myosin phosphatase target subunit 1(p-MYPT1)and autophagy-related proteins(LC3-I,LC3-II,p62)was detected by Western blotting.Results Compared with the normal group,pathological fibrosis of myocardial tissue was severe in the model group,HWI,content and area of collagen were increased,relative expression levels of type I procollagen,ROCK1 and TGF-β1 mRNA were increased,LC3-II/LC3-I was decreased,and expression level of p62 protein was increased(P<0.05).Compared with the model group,pathological fibrosis of myocardial tissue was mild in fasudil intervention group,HWI,content and area of collagen were decreased,relative expression levels of type I procollagen,ROCK1 and TGF-β1 mRNA were decreased,LC3-II/LC3-I was increased,and expression level of p62 protein was decreased(P<0.05).Conclusion T2MD can up-regulate expression of profibrotic cytokines in cardiomyocytes and inhibit cardiomyocyte autophagy to promote myocardial fibrosis by activating RhoA/ROCK pathway.Fasudil can interfere

关 键 词：2型糖尿病 心肌纤维化 法舒地尔 RhoA/ROCK通路 细胞自噬 

分 类 号：R587.1[医药卫生—内分泌]