温肺化纤汤动员自体骨髓间充质干细胞修复肺纤维化损伤的分子机制研究

There is still no effective treatment for pulmonary fibrosis until today. Our ten years clinical practice already proved Wenfeihuaxian formula has a good effect in patients with pulmonary fibrosis. The preliminary experimental study also showed that Wenfeihuaxian formula increased the migration ability of bone marrow mesenchymal stem cells in vitro. Autologous bone marrow mesenchymal stem cells characterized by self-renewal and multi-differentiation potentials can repair the damaged pulmonary tissue. The mechanism related to directional differentiation and regulation of local tissue cytokines. Therefore, We are interested in the effcet of Wenfeihuaxian formula mobilize autologous bone marrow mesenchymal stem cells to the lung injury site in vivo and its molecular mechanism. We plan firstly reconstitute the C57BL/6J mouse bone marrow, then built the model of bleomycinA5-induced pulmonary fibrosis, with different doses Wenfeihuaxian formula therapeutic intervention. The in vivo biological ability of Wenfeihuaxian formula mobilize and directional differentiation autologous bone marrow mesenchymal stem cells will be determined after 7,14,28 days. the molecular mechanism related to SDF-1/CXCR4 pathway will be fully studied. The completion of this project is not only an important guiding significance of traditional Chinese medicine basic research of pulmonary fibrosis. It is also expected to provide a safer, more effective treatment for many patients with pulmonary fibrosis.

目前治疗肺纤维化尚无十分有效的手段，我们十多年的临床实践证实温肺化纤汤对肺纤维化患者有较好的疗效，初步实验研究显示温肺化纤汤具有增加骨髓间充质干细胞体外迁移的能力。自体骨髓间充质干细胞具有自我更新及多向分化潜能，从骨髓中动员入血后可向受损的肺组织归巢并分化，并可通过释放和调节局部组织的细胞因子来达到修复的作用，因此温肺化纤汤是如何动员、定向募集自体骨髓间充质干细胞至肺损伤部位成为亟需解决的科学问题。本研究拟首先进行C57BL/6J小鼠的骨髓重建，继而采用博来霉素A5诱发肺纤维化模型，予不同剂量温肺化纤汤进行干预治疗，分7、14、28天3个时间点检测温肺化纤汤动员、定向募集自体骨髓间充质干细胞的体内生物学效能，同时以SDF-1/CXCR4通路为切入点，研究其分子机制。完成本项目不仅对肺纤维化的中医基础研究具有重要的指导意义，且有望为广大肺纤维化的患者提供更为安全、更为有效的新治疗手段。

温肺化纤汤临床治疗肺间质纤维化疗效明确，但是其作用机制不明晰。骨髓间充质干细胞从骨髓入血到达肺损伤处，具有良好的修复损伤作用。本项目对温肺化纤汤的组方原则、临床应用指征、中医护理措施进行总结；对其总黄酮、总多酚、总多糖、总皂苷含量进行测定；采用网络药理学技术对温肺化纤汤的关键成分群和靶点群进行筛选，并解析各关键成分之间的协同效应；对其抗炎、抗氧化能力进行系统评价；分离PMSC，观察温肺化纤汤对PMSC增值和迁移能力的促进作用，并在分子层面阐释其机制；构建博来霉素诱发肺间质纤维化大鼠模型，在整体动物模型层面证实温肺化纤汤抗肺间质纤维化与动员BMSCs迁移至肺损伤部位有关，上调SDF-1/CXCR4信号转导通路是其关键机制之一。本项目取得一批有价值的研究结果，发表相关论文19篇，有力地推动了温肺化纤汤的临床应用。