补阳还五汤干预Notch信号通路介导的内皮间质转化在特发性肺纤维化中的作用机制研究

Idiopathic pulmonary fibrosis (IPF) is a refractory disease manifested by aggravated and progressive dyspnea, restrictive ventilatory defects with reduced diffusion capacity, lung hypoxemia and diffuse radiographic lesions. Given that there is a lack of effective therapy for the disease, coupled with increased incidence in recent years, prevention and treatment for pulmonary fibrosis has become one of the major challenges and a hot spot in the medical field.Endothelial-to-mesenchymal transition (EndMT) is another currently found mechanism that influences the occurrence and development of IPF. Vascular endothelial cell is another important source of myofibroblast, and the transition is closely related with the Notch signal pathway. The interference of EndMT is of great significance to the prevention and inhibition of the occurrence of IPF.From the perspective of Traditional Chinese medicine, the deficiency of the lung and stagnation in collateral are the main pathogenesis of IPF, therefore the corresponding treatment should be supplementing qi and nourishing lung, as well as promoting blood circulation and dredging collaterals.Buyang Huanwu Tang not only embodies such therapeutic characteristics, but also has significant clinical effects to prevent and treat IPF. The research, focusing on the EndMT, is conducted to observe the influence Buyang Huanwu Tang has on the EndMT of IPF based on the IPF rat model and EndMT cell model induced in vitro; meanwhile, by adopting western blot, transfection, RNA interference and other methods to discuss the functional mechanism of Buyang Huanwu Tang intervening in the EndMT of IPF from the perspective of Notch signal pathway, for the purpose of seeking the key target of Buyang Huanwu Tang in the process of the IPF prevention and treatment.

特发性肺纤维化（IPF）是以进行性加重的呼吸困难、限制性通气障碍伴弥散功能降低、低氧血症以及双肺弥漫性病变为主要表现的难治性疾病，临床缺乏特效药物，是目前医学研究的热点。内皮间质转化（EndMT）是新近发现的影响IPF发生发展的又一重要机制，血管内皮细胞是肌成纤维细胞的又一重要来源，这一转化又与Notch信号通路密切相关。干预EndMT，对阻抑IPF的形成具有重要意义。中医学认为，肺虚络阻是IPF的主要病机，治当益气养肺，活血通络。补阳还五汤不仅体现了这一治疗特点，而且临床疗效显著，能够有效防治IPF。本研究围绕内皮间质转化为着眼点，从大鼠IPF动物模型和体外诱导EndMT细胞模型两方面观察补阳还五汤对IPF内皮间质转化的影响，并采用免疫印迹、转染、RNA干扰等多种方法从Notch信号通路探讨补阳还五汤干预IPF内皮间质转化的作用机制，从而寻求补阳还五汤防治IPF的关键靶点。

特发性肺纤维化（IPF）是一种病因未明，以进行性加重的呼吸困难、限制性通气障碍伴弥散功能降低、低氧血症以及双肺弥漫性病变为主要表现的难治性疾病，临床缺乏特效药物，是目前医学研究的热点。内皮间质转化（EndMT）是影响IPF发生发展的重要机制，血管内皮细胞是肌成纤维细胞的又一重要来源。干预EndMT，对阻抑IPF的形成具有重要意义。中医学认为，肺虚络阻是IPF的主要病机，治当益气养肺，活血通络。补阳还五汤不仅体现了这一特点，而且能够有效防治IPF。本项目以EndMT为着眼点，从大鼠IPF动物模型和体外诱导EndMT细胞模型两方面观察了补阳还五汤对IPF内皮间质转化的影响，并采用免疫印迹、转染、RNA干扰等多种方法从Notch信号通路探讨了补阳还五汤干预IPF内皮间质转化的作用机制。动物实验结果表明IPF大鼠肺组织存在EndMT，其内皮细胞标志物表达下调，而间质细胞标志物表达上调，补阳还五汤可通过干预EndMT来减轻大鼠的肺纤维化程度，其机制可能与调控TGF-β1/Smad3、Jagged1/Notch1、Dll4/Notch4信号传导有关。进一步体外细胞实验研究发现，TGF-β1可诱导HPAEC细胞发生间质转化，Notch信号通路参与了IPF的EndMT过程，阻断Notch信号通路可抑制EndMT，其中Notch受体Notch1和Notch4是HPAEC细胞发生间质转化的重要促进因素，补阳还五汤含药血清可通过调控Notch1和Notch4信号传导抑制内皮间质转化，从而防治IPF。本项目研究成果，表明了补阳还五汤可通过干预EndMT来防治IPF，其机制与调控Notch信号通路有关，同时也从实验角度证实了益气与活血配伍防治IPF具有协同增效的良好效果。这不仅为临床防治IPF提供了重要的理论指导和实验依据，也为经典名方补阳还五汤的二次开发及应用奠定了基础。