机械力作用诱导心脏瓣膜间质细胞TN-C表达及意义_医学论文.docVIP

机械力作用诱导心脏瓣膜间质细胞TN-C表达及意义_医学论文 机械力作用诱导心脏瓣膜间质细胞TN-C表达及意义_医学论文 作者：江磊 高洪波 徐平 李贞福 常青 郝传吉 王伟 于仁斌 【摘要】 目的 通过压力培养细胞装置研究主动脉瓣膜间质细胞在机械力作用下肌糖蛋白-C（TN-C）表达的改变，从瓣膜对机械力损伤修复角度探讨风湿性心脏瓣膜病的发病机制。方法 无菌手术获取主动脉瓣，剥除内皮层，胶原酶消化分离间质细胞，体外培养；构建产生等双轴周期性压力装置；利用机械力作用细胞后，以实时定量RT-PCR鉴定TN-C的表达；加入不同蛋白激酶阻断剂，分析参与机械力诱导TN-C的信号传导途径。结果 成功构建双轴周期性压力装置；机械力通过RhoA/ROCK途径诱导TN-C的转录，而p38MAPK、PKC和ERK途径没有参与机械力诱导TN-C的表达。结论 在风湿性瓣膜炎的发病过程中，机械力在TN-C的转录和表达中有一定作用，从而诱发瓣膜病理改变。 【关键词】 肌糖蛋白-C；机械力；风湿性心脏病；信号转导 　　［ABSTRACT］Objective［ To explore pathogenesis of rheumatic heart vascular diseases （RHVD） from direction of valve repair after mechanical stress injury through tenascin-C （TN-C） expression by culturing valve interstitial cells in vitro with designed strain-producing instrument under mechanical stress.［ Methods［ Aortic valves were obtained under aseptic condition, and the endothelial layer deprived. The valve interstitial cells were isolated by collagenase Ⅰ and then cultured in vitro. The equi-biaxial cyclic strain production device was developed. TN-C mRNA was measured by quantitative real-time RT-PCR after mechanical application. Signal transduction pathways induced by mechanical stress were assessed after using different protein kinase inhibitors.［ Results［ The equi-biaxial cyclic strain production device was developed successfully. Cyclic strain induced TN-C transcription through RhoA/ROCK signaling pathway other than p38 mitogen-activated protein kinase, PKC and ERK activation were found. ［ Conclusion［ In the process of rheumatic valvulitis, the mechanical power plays a certain role in TN-C transcription and expression and thus inducing pathologic changes of the valve. 　　［KEY WORDS］Tenascin-C Mechanical stress Rheumatic heart disease Signal transduction 风湿性心脏病（简称风心病）是危害人类健康的严重疾病，其发病机制并未完全确定。传统理论认为其与A型溶血性链球菌的感染和宿主自身免疫反应有关［1］。既往研究也发现，风湿性病变的发生部位主要集中在心脏瓣膜、心肌、关节等细胞外基质（ECM）成分承受机械力最大最频繁的部位，特别是风心病瓣膜病变的发生与承担的压力高低密切相关［2］。肌糖蛋白-C（TN-C）是ECM重塑的一组重要分子，且是迄今为止能够证实的由机械信号直接调控的最重要分子之一［3］。本研究旨在探讨机械力诱导TN-C在风心病瓣膜间质细胞中表达的机制。 　　1 材料和方法