基于TGF-β1/Smads信号传导通路的益气养阴活血通络法干预特发性肺纤维化机制研究

Idiopathic pulmonary fibrosis (IPF), the incidence and mortality rate of which is high,is difficult to treat in modern medicine. TCM treatment of it has embodies characteristics and advantages. Pre-clinical studies show the method of tonifying qi, nourishing yin, promoting blood circulation and dredging collaterals(Shenlong Decoction), based on the theory of "Lung deficiency with meridians stasis",has an effect on the treatment of IPF. Experimental studies have confirmed that Shenlong Decoction is able to slow down the process of lung fibrosis by anti-inflammatory and resistance to the imbalance of matrix production and degradation. With further research, more pathological links ,such as oxidative stress, Apoptosis and Th1/Th2 cytokines imbalance ,are being revealed,and the regulation of TGF-β1/Smads signal transduction pathways plays an important pivotal role in them.Our research group will take the study of TGF-β1/Smads signaling pathway as an entry point, conduct experiments of whole animal and cell culture, and use the methods of Western-blot、RT-PCR、flow cytometry、MTT and so on to clarify the mechanism that the method of tonifying qi, nourishing yin, promoting blood circulation and dredging collaterals is effective intervention in IPF by regulating multiple links of TGF-β1/Smads signaling pathways and then playing the role of antioxidant, anti-apoptotic and coordination Th1/Th2 cytokines balance. This will provide the scientific basis for prevention and treatment of IPF and lay a foundation for developing new drugs.

特发性肺纤维化（IPF）发病率、致死率较高，是现代医学面临的治疗难题，中医药对其治疗体现出特色和优势，前期临床表明，基于"肺虚络瘀"理论的益气养阴活血通络法（参龙煎剂）干预IPF有效；实验研究证明，参龙煎剂可通过抗炎、抗基质生成与降解失衡途径减缓肺纤维化进程。随着研究的深入，更多IPF的病理环节被揭示，如氧化应激、细胞凋亡、Th1/Th2型细胞因子失衡等，且发现TGF-β1/Smads信号通路调控失常在其中发挥重要的枢纽作用。本课题组拟开展整体动物实验及体外细胞实验，以TGF-β1/Smads信号通路研究为切入点，采用Western-blot、RT-PCR、流式细胞术、MTT等方法，阐明益气养阴活血通络法调控TGF-β1/Smads信号通路的多个环节，发挥抗氧化、抗细胞凋亡、协调Th1/Th2型细胞因子平衡作用，进而有效干预IPF的机制，为IPF的防治提供科学依据，并奠定新药研制的基础。

特发性肺纤维化（IPF）发病率、致死率较高，是现代医学面临的治疗难题，中医药对其治疗体现出特色和优势，前期临床表明，基于“肺虚络瘀”理论的益气养阴活血通络法（参龙煎剂）干预IPF有效；实验研究证明，参龙煎剂可通过抗炎、抗基质生成与降解失衡途径减缓肺纤维化进程。随着研究的深入，更多IPF的病理环节被揭示，如氧化应激、细胞凋亡、Th1/Th2型细胞因子失衡等，且发现TGF-β1/Smads信号通路调控失常在其中发挥重要的枢纽作用。本课题组拟开展体内及体外细胞实验，以TGF-β1/Smads信号通路研究为切入点，采用Western-blot、RT-PCR等方法，阐明益气养阴活血通络法调控TGF-β1/Smads信号通路的多个环节，发挥抗氧化、抗细胞凋亡、协调Th1/Th2型细胞因子平衡多方面的作用，进而有效干预IPF的机制。.实验结果：1.初步建立肺络构效失衡理论的IPF病、机、证、效体系；2. 益气养阴活血通络法能够有效抑制TGF-β1/Smads信号通路中发挥效应的TGF-β1、TGFβRII、Smad3蛋白及mRNA的高表达，促进发挥负性调控效应的Smad7蛋白/mRNA表达上调，推测其作用机制可能与TGF-β1/Smads信号通路有关；可能通过抑制IL-4蛋白和IL-4mRNA，增强INF-γ蛋白和INF-γmRNA表达以调解TH1/TH2平衡体系，进而延缓纤维化进展；可能通过调控TGF-β1/Smads信号通路，进而上调Bax、Fas、Fas L、CasPase-3含量和下调bcl-2含量促进肺成纤维细胞凋亡，从而延缓IPF进展；可能调控氧化应激相关氧自由基、抗氧化酶的作用，调节Keap1/Nrf2信号转导通路以及促进Keap1/Nrf2信号转导通路调控的下游靶蛋白和基因而实现的，进而延缓IPF进展。. 本实验阐明益气养阴活血通络法可能调控TGF-β1/Smads信号通路的多个环节抗细胞凋亡进而延缓IPF的作用机制，同时可能通过协调Th1/Th2型细胞因子平衡、抗氧化的作用多靶点、多环节减少或减缓成纤维化灶的形成、促进ECM的降解，延缓IPF进展，全面阐释益气养阴活血通络法干预IPF大鼠的可能作用机制，为IPF的防治提供科学依据，并为新药研制奠定基础。