益肺散结方负调控PI3K-AKT-mTOR通路激活自噬干预肺纤维化的分子机理研究

Pulmonary fibrosis, a terminal pathological change in the development of chronic lung disease, is pathological outcomes of abnormal tissue hyperplasia. Pulmonary fibrosis results in severe hypoxia and diffuse dysfunction, is a puzzle of clinical treatment. Our previous studies have confirmed that prescription of Yunnan traditional Chinese medicine "Yi Fei San Jie recipe" using Yi Qi, Zhu Yu and Di Tan policy could effectively improve the fibrosis of small airway, relieve airway remodeling in COPD. Based on the similarity between pulmonary fibrosis and small airway fibrosis, this study focuses on pulmonary fibrosis, and found that "Yi Fei San Jie recipe" could activate autophagy and significantly reduce the deposition of collagen, and relieve pulmonary fibrosis. Consequently, we will further study upstream pathway of PI3K-AKT-mTOR involving autophagy, regard imbalance in matrix degradation as the targets of Chinese medicine. We will verify the molecular mechanism of the recipe and the main active ingredient through the negative regulation of PI3K-AKT-mTOR pathway to activate autophagy, reduce collagen deposition and inflammatory factors, improve lung inflammation, lower substrate metabolism of lung microenvironment, and slow down the process of pulmonary fibrosis in the animal and cell models. The research will seek targets from the traditional Chinese medicine, and provide a theoretical basis for the treatment of pulmonary fibrosis.

肺纤维化是慢性肺疾病发展的终末病理改变，造成严重缺氧和弥散功能障碍，是临床治疗中的一个难题。我们前期研究证实，采用益气、逐瘀、涤痰策略的云南名中医验方“益肺散结方”能有效改善小气道纤维化，缓解COPD气道重塑发展。基于肺纤维化和肺小气道纤维化的病理相似性，本研究进一步聚焦肺纤维化，发现益肺散结方能够激活自噬基因表达，并显著降低胶原沉积、缓解肺纤维化，我们拟将该方调节纤维化的机制研究进一步深入到PI3K-AKT-mTOR自噬上游通路，以肺局部微环境中的基质降解失衡为中药作用靶点，从动物整体和细胞分子水平层面探讨该方及其主要活性成分通过负调控PI3K-AKT-mTOR通路激活自噬作用，减低胶原基因表达和炎症因子释放、下调肺微环境基质代谢，从而减缓肺纤维化进程的分子机制。为从中医药中寻求多靶点防治肺纤维化的发生发展提供理论依据。

肺纤维化是肺疾病发展的终末病理改变和结局，项目基于益肺散结方的疗效开展了相关机制研究，发现在肺纤维化小鼠模型组和益肺散结方的干预组中，通过Western Blot 检测LC3-II蛋白水平和电镜观察自噬小体数目，证明了益肺散结方会激活自噬。在肺纤维化模型组中，益肺散结方进行干预的同时，使用自噬抑制剂氯喹（CQ）处理，检测肺组织炎症程度和炎症因子表达、纤维沉积和肺纤维化相关基因的表达水平。阐明了自噬信号被抑制后影响了益肺散结方的药效。在细胞水平验证，益肺散结方中单味药的主要活性成分（黄芪总皂苷、莪术醇、姜黄素）负调控PI3K-AKT-mTOR激活自噬，影响炎症和胶原基因的分子机制。从动物整体和细胞分子水平层面揭示该方及其主要活性成分通过负调控PI3K-AKT-mTOR通路激活自噬下调肺微环境基质代谢，从而减缓肺纤维化进程的分子机制，为从中医药中寻求多靶点防治肺纤维化的发生发展提供理论依据。发表研究论文11篇，其中SCI论文6篇。培养研究生6人，其中中医基础理论研究生1人，中西医结合基础5人，4人继续读博。团队成员1人晋升二级教授，1人入选省级人才项目，1人晋升副高职称。