内皮间充质转化与急性病毒性心肌炎早期心肌纤维化的关系  

Relationship between endothelial-to-mesenchymal transition and cardiac fibrosis in acute viral myocarditis

作　　者：华军益[1,2] 张召才[3] 蒋旭宏[2] 何煜舟[2] 陈鹏[1] 

机构地区：[1]浙江大学医学院附属第一医院心内科,浙江杭州310003 [2]浙江中医药大学附属第一医院,浙江杭州310006 [3]浙江大学医学院附属第二医院心内科,浙江杭州310009

出　　处：《浙江大学学报（医学版）》2012年第3期298-304,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省自然科学基金(Y2080473);浙江省中医药科学研究基金(2011B532779)

摘　　要：目的:探讨心脏内皮间充质转化(EndMT)与急性病毒性心肌炎心肌纤维化形成的关系。方法:28只Balb/c小鼠随机分为对照组(n=8)、心肌炎组(n=10)和心肌炎+重组人骨形成蛋白7(rh-BMP7)干预组(干预组,n=10)。心肌炎组和干预组小鼠1次性腹腔接种柯萨奇病毒B3(CVB3),对照组小鼠腹腔注射等量病毒培养液,对照组和心肌炎组小鼠次日一次性腹腔注射生理盐水0.1 ml,干预组小鼠腹腔注射等量rh-BMP7(300μg/kg)以抑制转化生长因子(TGF-β1);7 d后处死小鼠取心脏,以苦味酸天狼星红染色后计算心脏胶原容积积分(CVF),实时RT-PCR法和Western blot方法检测小鼠心脏中TGF-β1、内皮细胞标志物(CD31和VE-cadherin)、成纤维细胞特异蛋白(FSP-1)、α-平滑肌肌动蛋白(α-SMA)和I型胶原(Col1α1)的基因和蛋白表达情况。结果:与对照组比较,心肌炎组小鼠心肌坏死区出现明显心肌纤维化,也出现了以内皮细胞表型(CD31、VE-cadherin)丢失、间充质细胞表型蛋白(FSP-1、α-SMA)和胶原Col1α1增多为特征的EndMT现象;同时,TGF-β1表达明显增高;抑制TGF-β1可以同时逆转EndMT和心肌纤维化。结论:EndMT参与急性病毒性心肌炎心肌纤维化的形成,TGF-β1是重要介导因子。Objective： To investigate the relationship between endothelial-to- mesenchymal transition （EndMT） and myocardial fibrosis in acute viral myocarditis （VMC）. Methods： Twenty-eight Balb/e mice were randomized into 3 groups：control group （n = 8 ）, VMC group （n =10） and intervention group （n =10）. Mice in VMC and intervention groups were injected intraperitoneally（i, p） with single dose of coxsackievirus B3, mice in control group were injected with equal amount of viral-free vehicle. In the following day, mice in control and VMC groups were injected i. p with 0.1 ml of saline and intervention group with 0.1 ml of recombinant human bone morphogenetic protein 7 （ rh-BMP7 ） at a concentration of 300 p,g/kg. The mice hearts were harvested after 7 d, cardiac collagen volume fraction （CVF） was calculated on picrosirius red-stained sections, mRNA and protein expression of TGF-β1, CD31, VE- cadherin,fibroblast special protein 1 （FSP-1） and u-smooth muscle aetin（α-SMA） and collagen 1α1 in myocardiac tissues were detected by real-time RT-PCR and Western blot analysis, respectively. Results： Compared to controls, overt fibrosis was presented in necrotic area of myoeardium in VMC group. Meanwhile,marked increase of TGF-~I expression accompanied with EndMT characterized by loss of endothelial phenotype （ decreased expression of CD31 and VE-cadherin） , gain of mesenchymal proteins （overexpression of FSP-1 and α-SMA） and increased synthesis of collagen was also demonstrated. Both EndMT and cardiac fibrosis were simultaneously reversed by TGF-β1 inhibition. Conclusion： EndMT is involved in cardiac fibrosis in acute viral myocarditis ,TGF-β1 might be a main mediator.

关 键 词：心肌炎/病毒学 纤维化/病理学 病毒性疾病 骨形态发生蛋白质类/药理学 转化生长因子Β 肠道病毒B型 人 

分 类 号：R542.21[医药卫生—心血管疾病]