中药复方与RNA干扰Gremlin基因表达对DN大鼠肾系膜细胞蛋白表达差异研究

In this study, preliminary studies on the basis of traditional Chinese medicine anddiabetic nephropathy to Jianpiyishenfang blood circulation disease closely related to theGremlin gene targeting, integrated use of protein chip, RNA interference, serum pharmacology, cell culture methods and techniques.By rat mesangial cells （RMCs）were cultured in the serum of Chinese herbal compound with highsugar or lentivirus-mediated Gremlin of RNAi and high-sugar , respectively, using the protein chipanalysis of traditional Chinese medicine the role of RMCs, Gremlin-RNAi lentivirusinfection RMCs ,GFP tracer slow the RMCs virus negative control, protein expression profile differences between the high sugar RMCs normal RMCs group of, to carry out thestudy of proteomics, analysis of the molecular mechanisms of diabetic nephropathy, in-depth understanding of the Gremlin's role in the pathogenesis of diabetic nephropathyand other complications, found determine the therapeutic targets of diabetic nephropathy, and deepen the study on the mechanism of prevention and treatment of diabetic nephropathy in Chinese medicine, led to the development of new highly effectivetraditional Chinese medicine preparation, and establish an efficient DN intervention programs, and explore new ways and means of integrative medicine, and thus for the development of new and efficient traditional Chinese medicine preparation foundation,are important.

本研究在前期研究的基础上，以健脾益肾活血化瘀中药复方和糖尿病肾病发病密切相关的Gremlin基因为对象，综合运用蛋白芯片、RNA干扰、血清药理学、细胞培养等方法和技术。 通过对高糖培养的大鼠肾系膜细胞（RMCs）分别加以中药复方血清和慢病毒介导Gremlin RNAi作用，采用蛋白芯片分析中药复方作用RMCs、Gremlin-RNAi慢病毒感染RMCs、GFP示踪慢病毒阴性对照RMCs、高糖RMCs和正常RMCs组间蛋白表达谱的差异，开展蛋白组学研究，分析糖尿病肾病的分子作用机制，对于深入了解Gremlin在糖尿病肾病等并发症发病过程中的作用，发现确定糖尿病肾病的治疗靶点，深化对中药防治糖尿病肾病作用机制的研究，进而开发新型高效中药制剂，建立高效的DN干预方案，探索中西医结合的新方法和途径，进而为开发新型高效中药制剂奠定基础，均有重要意义。

目的：本研究在前期研究的基础上，研究以健脾益肾活血化瘀中药复方和糖尿病肾病发病密切相关的Gremlin基因对RMCs蛋白质组表达的影响。方法：综合运用蛋白芯片、RNA干扰、血清药理学、细胞培养等方法和技术。通过对高糖培养的大鼠肾系膜细胞（RMCs）分别加以中药复方血清和慢病毒介导Gremlin RNAi作用，采用蛋白芯片分析中药复方作用RMCs、Gremlin-RNAi慢病毒感染RMCs、GFP示踪慢病毒阴性对照RMCs、高糖RMCs和正常RMCs组间蛋白表达谱的差异，开展蛋白组学研究。.结果：健脾益肾活血化瘀中药复方能显著改善RMCs GF-β1、VEGF、PDGF、CTGF、IL-6、MCP-1等蛋白的合成，减轻糖尿病肾病致病物质的累积，改善RMCs的功能。.结论：分析糖尿病肾病的分子作用机制，对于深入了解Gremlin在糖尿病肾病等并发症发病过程中的作用，发现确定糖尿病肾病的治疗靶点，深化对中药防治糖尿病肾病作用机制的研究，进而开发新型高效中药制剂，建立高效的DN干预方案，探索中西医结合的新方法和途径，均有重要意义。