基于调控PTEN/PI3K/AKT信号转导的补气活血通络方药治疗特发性肺间质纤维化机制

Idiopathic pulmonary fibrosis (IPF) is one of the most serious and complicated respiratory diseases, and is difficult to treat due to the lack of effective drugs. The pathogenesis of IPF has been linked to lung fibroblast cells, whose abnormal proliferation, migration and differentiation are considered as the primary disease-promoting factors. On the basis of confirmed curative effect of Traditional Chinese Medicine of invigorating qi and activating blood,dredging collaterals, the following study will be done:1.The rat model of IPF with bleomycin will be made. Rats will be divided into normal group, model group, prednisone group and invigorating qi and activating blood dredging collaterals herbs groups (to be respectively treated high, medium or low dosage). The impact of treatment on the PTEN/PI3K/AKT signaling pathway will be studied by examining the expression of PTEN, PI3K, p-AKT,et al in rat using Immunohistochemistry and Real-time PCR; 2.investigate the effect of treatment on the physiology of fibroblast cells (migration, proliferation and differentiation) and gene expression changes using in vitro cell culture, serum pharmacology and molecular biology techniques. Differential expression of the key genes and proteins in the PTEN/PI3K/AKT pathways will be studied. The significance of the study is to provide evidence for the clinical application of Traditional Chinese Medicine in IPF treatment and prevention, and to understand the mechanisms of invigorating qi and activating blood,dredging collaterals herbs treatment in the IPF pathogenesis.

特发性肺间质纤维化（IPF）是呼吸系统疑难危重病之一，临床缺乏有效治疗药物，肺成纤维细胞（FB）增殖迁移、分化是促肺纤维化关键因素。基于前期实验研究基础及确证的补气活血通络方药治疗IPF的临床疗效，本项目拟研究内容为：一以博来霉素制备IPF大鼠模型，设空白组、模型组、泼尼松组及补气活血通络方药高、中、低剂量组，从肺功能、肺组织病理学观察方药疗效；通过免疫组化法及实时荧光定量PCR法检测肺组织中PTEN、PI3K、p-AKT等表达，观察方药对PTEN/PI3K/AKT信号通路指标的影响；二采用细胞培养、血清药理学和分子生物学等技术，观察大鼠FB的增殖迁移及分化，测定PTEN/PI3K/AKT信号通路中关键基因与蛋白表达。通过研究，评价补气活血通络方药对FB增殖迁移、分化的影响，基于调控FB细胞PTEN/PI3K/AKT信号通路，阐释补气活血通络方药治疗IPF作用机制，为临床应用提供科学依据。

本课题拟研究补气活血通络方药（BHTF）治疗特发性肺间质纤维化（IPF）可能的作用机制，一方面通过气管插管注入博来霉素法复制IPF大鼠模型，观察肺功能、肺组织病理学及PTEN、PI3K、p-AKT等指标的表达；另一方面体外培养肺成纤维细胞，采用细胞培养、血清药理学和分子生物学等技术，观察大鼠成纤维细胞（FB）的增殖迁移及分化，测定PTEN/PI3K/AKT信号通路中关键基因与蛋白表达。通过BHTF对PTEN/PI3K/AKT信号通路指标的干预研究，探讨BHTF对IPF的治疗作用，为临床提供依据。研究结果显示：1、采用气管插管注入博来霉素法可成功复制IPF大鼠模型；2、与模型组及泼尼松组比较，BHTF组能减轻大鼠肺组织肺泡炎症改变及纤维组织增殖，控制大鼠肺功能的恶化，尤其调节通气功能、肺组织顺应性、肺容量及稳定肺泡表面张力疗效显著；3、BHTF组较模型组及泼尼松组能上调PTEN水平，下调TGF-β1、PI3K、AKT、mTOR的蛋白和基因表达。结果表明，补气活血通络方药能有效改善IPF大鼠肺功能、抑制肺组织纤维化增生程度，上调PTEN水平，下调TGF-β1、PI3K、AKT、mTOＲ、α-SMA指标，揭示补气活血通络方药治疗IPF可能存在的作用机制，为临床提供一定依据。已发表中文核心4篇，英文SCI收录1篇（IF2.06），协助培养硕士研究生1名。