基于MTC技术探讨整合素αvβ3力学传导调控VICs钙化机制研究

Calcified heart valvular disease(CHVD) is a common disease, causing serious harm to human health. VICs is the key point of the calcification of valves, and abnormal mechanical stimulation is an important initial factor of it. However, the mechanism is not clear. We have founded that MTC can apply accurate mechanical stimulation in microenvironment to certain cells to explore the force transmission between cells and ECM. We have also found that calcified valves express more integrin αvβ3. Besides, integrin αvβ3 plays a role in the force transmission inducing the change of the calcified genes of VICs. Based on the preliminary results, we will use MTC which has proprietary intellectual property rights to mimic force transmission between ECM and VICs and explore the role of the integrin αvβ3 in the force transmission and promoting calcification of valves.  Finally, we will combine MTC with siRNA gene silencing and plasmids encoding fluorescent proteins to try to explore the mechanism in the αvβ3 activating causing the change of calcified genes of VICs and the molecular networks mechanisms of mechanical stimulation-integrin αvβ3 activating- force transmission-calcification of VICs. Besides, we hope to provide the theoretical foundation for finding the potential target of calcification.

钙化性心脏瓣膜病是常见多发病，严重危害人类健康。瓣膜间质细胞(VICs)是瓣膜钙化的核心环节，异常力学刺激是导致VICs钙化的重要始动因素，但其机制不明。课题组前期研究表明：①磁力扭曲仪(MTC)可通过精准调控微观力学刺激应用于特定单个细胞与细胞外基质(ECM)力学传导研究; ②钙化瓣膜中VICs表面整合素αvβ3表达增加；③整合素αvβ3在MTC模型中通过传导力学信号快速诱导VICs细胞核内钙化基因变化。基于此，本项目拟利用独立自主知识产权的MTC技术，模拟ECM与VICs间力学传导过程，明确整合素αvβ3参与力学传导并促进瓣膜钙化的作用；进一步结合siRNA基因沉默及荧光质粒技术，特异性精准施加干预措施，探讨整合素αvβ3激活调控VICs钙化基因变化的机理，阐明力学刺激-整合素αvβ3激活-力学信号传导-VICs钙化发生发展机制，为寻找潜在特异性抗钙化靶点提供理论依据。

钙化性心脏瓣膜病是常见多发病，严重危害人类健康。瓣膜间质细胞(VICs)是瓣膜钙化的核心环节，异常力学刺激是导致VICs钙化的重要始动因素。课题组研究发现磁力扭曲仪(MTC)可通过精准调控微观力学刺激应用于特定单个细胞与细胞外基质(ECM)力学传导，并可刺激瓣膜间质细胞成骨样分化。基于此，本项目以独立自主知识产权的MTC技术为基础，模拟ECM与VICs间力学传导过程，1）构建MTC模型，研究微观力学刺激作用大小，作用时间，作用频率，得到微观力学最佳促钙化参数。2）深入研究力学传导过程，发现力学受体整合素αvβ3受体在MTC模型中可快速诱导VICs成骨样分化。3）深入研究αvβ3受体参与信号通路，通过RGD结构域与整合素αvβ3结合，发现其可通过激活MAPK信号通路，然后p-ERK的易位激活成骨基因的表达，从而诱导VICs成骨样分化。本项目明确整合素αvβ3参与力学传导并促进瓣膜钙化的作用；探讨整合素αvβ3激活调控VICs钙化基因变化的机理，阐明力学刺激-整合素αvβ3受体-MAPK信号传导-VICs钙化发生发展机制，为寻找潜在特异性抗钙化靶点提供理论依据。