基于Ezrin蛋白的针刺抗哮喘舒张气道平滑肌生物学机制研究

本项目建立在临床有效的针刺抗哮喘方法和生物学机制研究有所发现的基础上。利用基因组学技术发现金属硫蛋白2(MT-2)是重要的针刺抗哮喘效应因子，可通过膜受体肌动蛋白结合蛋白2(TG2)舒张气道平滑肌。然而TG2介导的气道平滑肌收缩舒张分子机制尚不清楚。利用蛋白磷酸化芯片发现TG2激活后可显著增强气道平滑肌细胞Ezrin蛋白磷酸化，经免疫共沉淀和表面等离子共振技术证实TG2和Ezrin可形成稳定蛋白复合体。我们推测Ezrin可结合TG2受体并在下游信号传导通路中起关键作用。本项目拟研究针刺对Ezrin表达、磷酸化影响，检测Ezrin在哮喘呼吸功能、气道平滑肌细胞刚度、牵张力、收缩舒张等生物学作用和针刺调节效应，阐明Ezrin蛋白在TG2受体介导的气道平滑肌收缩舒张调节机制中作用，以期为针刺抗哮喘效应生物学内涵阐释提供新的科学依据。

This project is based on a clinically proven method of acupuncture for treating asthma and the previous achievement in biological mechanism research. Previous studies have shown that metallothionein 2 (MT-2) was one of the most important anti-asthmatic factors of acupuncture and it relaxed airway smooth muscle via transgelin-2 (TG2) receptor. However, the molecular mechanism of contraction and relaxation of airway smooth muscle by TG2 receptor is still not clear. It was found that MT-2 significantly increased the level of ezrin phosphorylation via TG2 receptor by using a protein antibody array. The strong interaction between ezrin and TG2 was subsequently confirmed by co-immunoprecipitation and surface plasmon resonance. Therefore, ezrin was thought to play a key role in the transduction of TG2 receptor signaling pathway. This project intends to study the effect of acupuncture on ezrin expression and phosphorylation, to observe the effect of ezrin on the respiratory function, the cellular stiffness, the traction force and relaxation of airway smooth muscle and the corresponding acupuncture regulation. This study will identify the molecular mechanism of ezrin in contraction and relaxation of airway smooth muscle mediated by TG2 receptor, which may provide a new scientific basis for the biological mechanism of acupuncture in treating asthma.