基于PI3K/AKT/HIF-1α通路的益气养阴活血法干预特发性肺纤维化血管新生“开关”机制研究

Idiopathic pulmonary fibrosis (IPF) is a disease with high mortality, rapid progress, pathogenesis is not clear, and Western medicine has no effective treatment of invasive fibrogenic proliferative disease. In recent years, it has been found that vascular hyperplasia is an important factor in the development of idiopathic pulmonary fibrosis. The activation of PI3K / AKT / HIF-1α signaling pathway is closely related to angiogenesis, A new target for prevention and treatment of pulmonary fibrosis. According to the characteristics of IPF TCM syndromes proposed "lung disease treatment, due to virtual stasis" theory, the occurrence of IPF to lung and kidney qi and yang deficiency, blood stasis lung line as the standard, there is lung disease, From the gas to the blood, lung stasis of the process, according to the proposed Qi and Yin and blood circulation treatment. Previous studies have shown that Yiqi Yangyinhuoxue method can improve the immune function of patients, reduce the degree of pulmonary fibrosis. Based on the previous study, the method of PI3K / AKT / HIF-1α signaling pathway was used as the starting point, and the patients were treated with ELISA, Western blot and RTqPCR. The relationship between the imbalance of vascular neonatal switch and idiopathic pulmonary fibrosis and the mechanism of the intervention of qi and yin and promoting blood circulation to provide new ideas for the prevention and treatment of idiopathic pulmonary fibrosis.

特发性肺纤维化（IPF）是一种死亡率高，进展快，发病机制尚不明确，且西医尚无有效治疗措施的隐袭性纤维增生性疾病。近年来研究发现血管新生“开关”失衡导致的血管过度新生是推进IPF发展的重要因素，而PI3K/AKT/HIF-1α信号通路的活化与血管新生密切相关，阻断血管新生是IPF防治的新靶点。我们据IPF中医证候学特点提出“肺失治节、因虚致瘀”论，IPF的发生以肺肾气阴两虚为本，瘀血阻滞肺络为标，存在肺失治节、由气到血，肺络瘀滞的过程，据此提出益气养阴活血的治法。既往研究表明，益气养阴活血法能延缓IPF的进程。本项目拟在前期研究基础上，采用体内外实验结合的方法，运用Western blot、RTqPCR等分子生物学技术，以PI3K/AKT/HIF-1α信号通路为切入点，探究“肺失治节，因虚致瘀”与IPF血管新生“开关”失衡的相关性及益气养阴活血法干预的分子机制，为IPF的中医药防治提供依据。

特发性肺纤维化（IPF）是一种死亡率高，进展快，发病机制尚不明确的隐袭性纤维增生性疾病。本项目在前期研究基础上，采用体内外实验结合的方法，运用Western blot、RTqPCR等分子生物学技术，以PI3K/AKT/HIF-1α信号通路为切入点，探究“肺失治节，因虚致瘀”与IPF血管新生“开关”失衡的相关性及益气养阴活血法干预的分子机制。.体内实验结果显示：IPF大鼠存在血管新生“开关”失灵，促血管新生因子HIF-1α、PDGF和血管新生抑制因子PEDF、ES动态平衡被破坏，出现异常的血管新生。模型组肺组织中PI3K、AKT、HIF-1α及VEGF的平均蛋白及mRNA表达量均高于空白组（P＜0.05），说明随着IPF的发生发展PI3K/AKT/HIF-1α信号通路被激活，从而调控VEGF的表达来影响血管的生成。而益气养阴活血法组方的参七虫草组大鼠肺组织中PI3K、AKT、HIF-1α及VEGF的平均蛋白及mRNA表达量均较模型组低（P＜0.05），推测参七虫草方可能通过抑制VEGF的表达抗肺纤维化。.体外实验结果显示：IPF模型组大鼠肺微血管内皮细胞迁移细胞数显著增多较空白组（P＜0.01），10%含药血清组、LY294002组及LY294002+10%含药血清组的迁移细胞数显著减少(P＜0.01)。模型组大鼠肺微血管内皮细胞中VEGF、PI3K、AKT、HIF-1α及mRNA表达水平均显著升高（P＜0.01，P＜0.05），10%含药血清组及LY294002+10%含药血清组VEGF、PI3K、AKT、HIF-1α及mRNA表达显著下降（P＜0.01，P＜0.05）。表明10%参七虫草含药血清对肺微血管内皮细胞的增殖抑制作用最强，血管新生参与IPF的发病过程，益气养阴活血法组方的参七虫草方可以通过抑制肺微血管内皮细胞迁移及VEGF蛋白及mRNA表达及抑制PI3K/AKT/HIF-1α信号通路，一定程度上干预肺纤维化形成。.研究发现血管新生“开关”失衡导致的血管过度新生是推进IPF发展的重要因素，而PI3K/AKT/HIF-1α信号通路的活化与血管新生密切相关，阻断血管新生是IPF防治的新靶点,研究结果验证了IPF“肺失治节，因虚致瘀”理论，益气养阴活血法可能通过PI3K/AKT/HIF-1α信号通路抑制VEGF的表达来干预血管新生，从而发挥对IPF的治疗效果。