ATRA通过ERK/MAPK通路调控AS兔心脏MLCK的表达  

Regulation of all-trans-retinoic acid on expression of myosin light chain kinase in myocardial tissue through the pathway of ERK/MAPK in atherosclerosis model rabbit

作　　者：王雪[1] 余沛 王怡[1] 周青[1] 汪渊[1] 朱华庆[1] Wang Xue;Yu Pei;Wang Yi(Dept of Biochemistry and Laboratory of Molecular Biology of Anhui Medical University, Key Laboratory of Gene Utilization for Severe Disease of Anhui Province,Hefei 230032)

机构地区：[1]安徽医科大学分子生物学实验室,安徽省/省部共建教育部重要遗传病基因资源利用重点实验室,合肥230032

出　　处：《安徽医科大学学报》2018年第10期1494-1499,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号：81570419)

摘　　要：目的研究全反式维甲酸(ATRA)通过信号调节激酶/丝裂原活化蛋白激酶(ERK/MAPK)通路调控动脉粥样硬化(AS)模型兔心脏中肌球蛋白轻链激酶(MLCK)的表达。方法普通新西兰大耳朵白兔随机分为正常组、模型组、ATRA组,正常组给予普通饲料喂养,模型组和ATRA组给予高脂饲料(普通饲料+1%胆固醇+5%猪油)喂养,并且ATRA组同时给予ATRA灌胃5 mg/(kg·d)。12周后,取动脉和心脏组织,油红染色分析动脉壁斑块形成情况;HE染色和Masson染色观察心肌组织形态学变化,免疫组织化学染色观察心脏中MLCK的表达;Western blot法检测各组MLCK蛋白和ERK蛋白磷酸化水平。体外培养H9c2心肌细胞,分为正常组、模型组、ATRA组、ERK/MAPK选择性抑制剂(PD)组。Western blot法检测各组MLCK蛋白的表达水平。结果 12周以后,AS兔模型建立成功。与正常组比较,模型组动脉壁形成大量粥样斑块;HE染色结果显示模型组心肌组织出现严重的紊乱情况;Masson染色结果显示模型组心肌组织发生明显胶原纤维堆积;免疫组化结果显示模型组心肌组织中MLCK表达量增多;而给予ATRA以后,心肌组织紊乱情况得到改善,胶原纤维堆积得到缓解,MLCK表达量和ERK磷酸化水平均有所降低。并且模型组细胞中MLCK蛋白表达升高,给予ATRA和PD98059后,MLCK表达降低。结论 ATRA可能通过ERK/MAPK通路降低模型兔心肌细胞MLCK的表达。Objective To explore the expression of myosin light chain kinase(MLCK)in myocardial tissue regulated by the all-trans-retinoic acid(ATRA)through the pathway of ERK/MAPK in atherosclerosis(AS)model rabbit.Methods The New Zealand white rabbits were randomly divided into three groups:normal group,model group and ATRA group.The normal group were fed with normal diets.The model group were fed on high-fat diets(normal diets+1%cholesterol+5%lard).The ATRA group was given ATRA[5 mg/(kg·d)]while feeding high-fat diets.After 12 weeks,the arterial and cardiac tissues were taken and the formation of arterial wall plaque was analyzed by oil red staining;Haematoxylin-eosin(HE)staining and Masson staining were used to detect morphological changes of myocardial tissue.Immunohistochemistry was used to detect the expression of MLCK in myocardial tissue.Western blot was used to observe the changes of MLCK and ERK phosphorylation in myocardial tis-sue.H9c2 cardiomyocytes were cultured in vitro and divided into normal group,model group,ATRA group,ERK/MAPK selective inhibitor(PD)group.Western blot was employed to detect the expression of MLCK.Results The AS model rabbit was built successfully 12 weeks later.Compared with normal group,arterial wall formed lots of atherosclerotic plaques in model group;HE staining showed that there was a serious disturbance of myocardial tissue in the model group;Masson stain indicated that collagen fibers accumulated seriously in myocardial tissue.Immunohistochemistry showed that expression of MLCK and ERK phosphorylation were enhanced in myocardial tissue.After the treatment of ATRA,the expression of the MLCK and ERK phosphorylation decreased clearly.The expression of MLCK protein in the cells of the model group was increased,and the expression of MLCK decreased after the ATRA and ERK/MAPK inhibitors were given.Conclusion ATRA may regulate the expression of MLCK in myocardial tissue of AS model rabbit by the pathway of ERK/MAPK.

关 键 词：全反式维甲酸 动脉粥样硬化 心肌组织 肌球蛋白轻链激酶 丝裂原活化蛋白激酶 

分 类 号：R541.4[医药卫生—心血管疾病] R972.6[医药卫生—内科学]