基于mTOR/SIRT1信号通路的补肺益肾方调控肺泡上皮细胞治疗COPD机制研究

慢性阻塞性肺疾病（COPD）严重危害公众健康，目前缺乏理想的治疗方法。细胞凋亡和自噬是COPD的重要病理生理机制，mTOR/SIRT1信号通路在此过程中起着重要作用，基于以往研究，进一步探讨补肺益肾方治疗COPD的作用机制，拟开展：①制备大鼠COPD稳定期模型，以阻断剂雷帕霉素为对照，从肺功能，外周血、肺泡灌洗液细胞因子表达，肺组织病理和超微结构观察补肺益肾方治疗COPD的疗效；采用免疫组织化学、TUNEL、qPCR、WB技术，从肺组织细胞凋亡、自噬及mTOR通路相关基因和蛋白表达，探讨补肺益肾方治疗COPD的作用机制。②以肺泡上皮细胞为研究对象，以雷帕霉素和Sirtinol为阻断剂，采用细胞培养、分子生物学等技术，围绕mTOR信号通路，观察补肺益肾方含药血清对香烟烟雾刺激下肺泡上皮细胞分泌细胞因子及细胞凋亡和自噬等影响。以揭示中医药治疗COPD作用机制，为中医临床应用和新药研发提供依据。

Chronic obstructive pulmonary disease (COPD) is a major public health problem worldwide and results in an economic and social burden, and without appropriate pharmacologic therapy. Cell apoptosis and autophagy are one of the principal physiopathologic mechanism of COPD. There is increasing evidence that signalling though the mammalian target of rapamycin (mTOR) / Sirtuin type 1（SIRT1） pathway is of critical importance in cellular senescence and ageing, It is reported that treatment with traditional Chinese medicine (TCM) may do some benefit to the disease. .On the basis of previous research, The aim of the study is to investigate the cellular and molecular mechanisms of TCM. Our work focuses on two parts: ①we preparing animal models of stable COPD, taking rapamycin as the inhibitors, evaluating the therapeutic effect of TCM on animal models by pulmonary function, cytokine expression in blood and BALF, pathology and ultrastructure. By way of immunohistochemistry, TUNEL、qPCR and western blot method, investigating mechanism of Bufei Yishen decoction on COPD by evaluating lung tissue apoptosis and autophagy and related gene and protein expression of mTOR/ SIRT1 pathway.②Culturing of alveolar epithelial cells A549, and preparing containingTCM medicine serum. By activated of cigarette smoke extract and inhibited by rapamycin and sirtinol, the study is to investigate the secretion of cytokines of A549 cells and cell apoptosis and autophagy, and the serum was used to do inhibitory tests in vitro. The related gene and protein of mTOR/SIRT1 pathway were detected by molecular biotechnology such as q-PCR、western blot method. The experiment would investigate the related mechanism of TCM on COPD, and would provide scientific evidence for research of new drugs and TCM clinical application.