Wnt/β-Catenin信号通路在气虚血瘀型特发性肺间质纤维化大鼠的动态表达及益气活血法的干预研究

Idiopathic pulmonary fibrosis (IPF) is a kind of unknown cause and poor prognosis respiratory system disease.At present, the pathogenesis is not very clear and no effective treatment. TCM Clinical medicine has confirmed its evolution of the pathogenesis is deficiency of Qi and blood stasis, Qi and activating blood circulation method can improve the clinical symptom and prognosis for IPF patients, but the mechanism is still not clearly. Previous studies have shown that Wnt/β-catenin signaling pathway plays an important role in the pathogenesis of IPF. Based on The pathogenesis of IPF and the existing research results, This study will use immunohistochemistry, Western blot, RT-PCR technology and histopathology methods to observe the Dynamic expression of Wnt/β-catenin signaling pathway and its downstream target cytokine including connective tissue growth factor, hepatocyte growth factor, matrix metalloproteinase and α-SMA, HYP, collagen type I, III collagen’s levels and tanalyze the correlation between the above index and the expression level on Wnt/β -catenin signal pathway. On the basis of the above research we choose the supplementing Qi and activating blood circulation method‘s representative prescription - “yang-supplementing and half-recovering decoction” to intervene in IPF rats. Through the observation about the Corresponding relationship of Wnt/β -catenin signaling pathway and its downstream target cytokine, we hope that through the above experiment will revealled mechanism of “ yang-supplementing and half-recovering decoction” in prevention and treatment of qi deficiency and blood stasis syndrome type IPF. To provide a theoretical basis for the prevention and treatment of IPF with traditional Chinese medicine.

特发性肺间质纤维化（IPF）是一种原因不明、预后不良的呼吸系统疾病，目前无有效的治疗方法。中医临床证实其病机演变多为气虚血瘀，以益气活血法治疗IPF可改善临床症状及预后，但有关该治法指导下的中药复方防治IPF的作用机制尚不明确。研究表明Wnt/β-catenin信号通路在IPF的发病过程中起重要作用。基于IPF的病机规律及现有研究基础，本研究采用免疫组化、免疫印迹、RT-PCR技术结合病理组织学，观察Wnt/β-catenin信号通路及其下游靶基因细胞因子CTGF、HGF、MMP-7的动态表达与α-SMA、HYP、Ⅰ型胶原、Ⅲ型胶原的相关性；在此基础上采用益气活血代表方补阳还五汤进行干预，从Wnt/β-catenin信号通路与下游细胞因子的对话效应揭示补阳还五汤防治气虚血瘀型IPF的作用机制，为中医药防治IPF提供理论依据。

特发性肺间质纤维化（idiopathic pulmonary fibrosis，IPF）是一种原因不明，进行性致纤维化的间质性肺炎，是一种慢性进行性不可逆转也是最常见的一种致命性肺疾病，目前无有效的治疗方法。中医临床证实其病机演变多为气虚血瘀，以益气活血法治疗IPF可改善临床症状及预后，多年来，我们在临床工作中观察到补阳还五汤在干预IPF中显示出较好的疗效。研究表明Wnt/β-catenin信号通路在IPF的发病过程中起重要作用。据此我们提出“补阳还五汤抗肺纤维化的作用机制是否与其调控Wnt/β-catenin信号通路有关”的假说，进一步研究益气活血法对 IPF 的干预作用，本研究采用免疫印迹、RT-PCR及ELISA结合病理组织学等观察Wnt/β-catenin信号通路在气虚血瘀型IPF大鼠的动态表达；益气活血法对气虚血瘀型IPF大鼠整体状态及肺组织形态学的影响；益气活血法对气虚血瘀型IPF大鼠wnt/β-catenin信号通路的影响。取得了重要的研究成果：①成功复制了气虚血瘀型IPF大鼠模型，并通过证候形态学评分、肺组织HE、Masson染色验证建模成功；并发现Wnt/β-catenin信号通路中Wnt1、β-catenin和下游靶基因MMP-7以及CTGF、α-SMA、HGF在IPF大鼠模型中表达显著升高且呈现动态表达；②补阳还五汤显著改善气虚血瘀型大鼠IPF肺组织颜色、外观、硬度、弹性，并显著减轻了大鼠肺部炎症及纤维化程度；③补阳还五汤能有效下调Wnt/β-catenin信号通路中Wnt1、β-catenin及下游靶基因MMP-7的表达，并调控CTGF、HGF、α-SMA、I型胶原、III型胶原的表达。从分子水平揭示中医益气活血法与抗纤维化治疗的关系，为中医药防治 IPF 提供理论及实验依据。