基于整合素/ILK/PI3K细胞信号通路探讨加味当归补血汤对慢性肾脏病治疗的机制

Basic on the research which we have finished, it verified that the way of tonifying kidney and blood circulation by Chinese herbs can protect the function of podocyte and reduce the renal tissue fibrosis. We analyze the reason of how to let the lesion of renal filtration membrane which including endothelial cell (podocyte), endothelial cell (EC) and glomerular basement membrane (GBM). We analyze the relationship between the degree of the lesion of renal filtration membrane and the extent of the urine protein too. .We find that the cell signals transduce may be the real reason which affect the intact of renal filtration membrane and the extent of the proteinuria. The cell signal pathway includes integrins link kinase (ILK), phosphoinositides 3-kinase (PI3K) and protein kinase B (PKB) and so on. We believe that the cell signal pathway can regulate the level of cell function, the gene for cell cycle and metabolic enzyme to extracellular matrix (ECM)..First, we want to know the relationship between the lesion of renal filtration membrane and the changing of cell signal pathway through doing research. Second, we want to know the changing of cell signal pathway both in vivo and in vitro through doing research. Third, we want to verify the Chinese herbs can decrease the proteinuria and protect the renal filtration membrane through regulating the cell signal pathway. We set up the blueprint to do the research through regulating the cell signal transduce to provent the development of kidney disease. We will use some advanced biology technology which including immunofluorescent technique, western blot and isotopic tracer technique and so on.

已有工作证实益肾通络法中药方剂对保护肾小球足细胞、抑制肾间质纤维化疗效确切。肾小球滤过屏障包括足细胞、内皮细胞和基底膜三部分。本研究拟从肾小球滤过屏障病变是蛋白尿产生的根本原因出发。结合以往研究基础和文献综述，我们认为细胞间信号传导可能是导致肾小球病变蛋白尿的核心机制，拟以"调控细胞间信号转导，改善肾脏病变进展"的视角，探讨益肾通络法中药方剂"加味当归补血汤"对慢性肾脏病治疗上的有效性。通过体内外动物实验如阿霉素肾病模型、足细胞、内皮细胞体外单培养和共培养观察其不同状态下"ILK/PI3K/PKB"等信号转导通路以及其对相关细胞因子、细胞周期基因和细胞外基质代谢相关酶的调控作用对肾小球滤过屏障的影响并分析其机制。实验拟采用蛋白质印记技术、免疫荧光技术和同位素示踪技术等先进的生物学手段，分析中药方剂加味当归补血汤作用的分子机制，为中医药防治慢性肾脏病提供科学依据。

本课题的研究工作按照国家自然科学基金申报书的工作安排主要以下几个方面的研究工作：1.进行整合素/ILK与ILK/PKB/PKC信号转导通路、细胞周期调节的P53基因和FAS蛋白表达与SD功能蛋白分子表达情况的相关实验。2. 进行应用中药方剂及其组分在实验动物模型的药效学研究，筛选最佳的中药组分配伍及活性组分配比。3.进行中药方剂活性成分在延缓肾脏纤维化，保护慢性肾脏病模型大鼠肾脏功能的研究。4.利用中药制药工程学的方式开展中药复方及其活性成分新制剂的开发研究工作并取得了一定的进展。相关研究工作证实了加味当归补血汤在慢性肾脏病中的治疗作用和保护机制。课题所用方剂获得国家发明专利授权保护。研究方剂在得到研究单位伦理委员会批准后开展临床研究工作验证疗效并进行成果转化工作。相关研究获得2015年度国家中医临床研究基地业务建设第二批科研专项（JDZX2015096）的资助。. 研究工作开展四年以来，按照工作进度开展工作顺利，先后申报发明专利5项，授权发明专利2项；发表课题相关SCI论文7篇，EI论文1篇，中文核心期刊16篇；培养研究生6名；课题负责人被推选为中国中西医结合学会青年工作委员会副主任委员，世界中医药联合会肾脏病委员会理事，中国中医药促进会肾脏病专业委员会理事；课题工作获得江苏省中医药科技进步三等奖；课题相关的研究工作顺利完成。