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目的观察中药复方肾消康对糖尿病肾病(DN)大鼠的防治作用及对肾脏基因表达的影响,探讨糖尿病肾病的发病机制,揭示中药复方肾消康防治DN的作用机理,筛选药效作用靶点,为临床推广应用提供科学依据。方法实验采用链脲佐菌素(STZ)加高热量饮食建立糖尿病大鼠肾脏损伤模型。运用放免、生化、光镜、电镜及美国Affymetrix公司的基因表达谱芯片、RT-PCR等先进检测手段和方法,观察了多项指标,尤其是通过聚类分析寻找和DN有重要相关性的基因,并进一步采用实时荧光定量RT-PCR法做验证实验研究。结果 S100钙结合蛋白A9(S100A9),在糖尿病大鼠肾脏表达上调,肾消康治疗组表达下调。结论应用Affymetrix Rat2302.0基因表达谱芯片检测糖尿病大鼠肾脏基因的表达,S100A9是筛选出的肾脏差异表达基因和药效靶点之一,并对该基因做荧光定量RT-PCR测序分析,这在国内外尚属首次。基因功能分析表明,在糖尿病状态下,S100A9表达上调,与糖尿病肾脏损伤有重要相关性,而经肾消康治疗后大鼠肾脏组织中S100A9表达下调,可见肾消康对S100A9基因表达具有良性调节作用,其作用机制还有待于进一步研究。
Objective To observe the preventive and therapeutic effects of traditional Chinese medicine Shenji Decoction on diabetic nephropathy (DN) rats and its effect on kidney gene expression, explore the pathogenesis of diabetic nephropathy and reveal the mechanism of traditional Chinese medicine Shenji Decoction in preventing and treating DN, and screen the pharmacodynamic target Point, to provide a scientific basis for clinical application. Methods The model of renal injury in diabetic rats was established by using streptozotocin (STZ) plus high-calorie diet. A number of indicators were observed using radioimmunoassay, biochemistry, light microscopy, electron microscopy and gene expression profiling chips from Affymetrix in the United States, and other advanced detection methods and methods, especially by cluster analysis to find genes with important relevance to DN , And further use of real-time fluorescence quantitative RT-PCR method to do validation experiments. Results S100 calcium binding protein A9 (S100A9) was up-regulated in the kidney of diabetic rats and down-regulated in the Shenkukang treatment group. Conclusion Affymetrix Rat2302.0 gene expression microarray was used to detect the expression of renal gene in diabetic rats. S100A9 was one of the differentially expressed and pharmacologically active targets of kidney, and the gene was analyzed by RT-PCR. For the first time at home and abroad. Analysis of gene function showed that S100A9 expression was up-regulated in diabetes, which was correlated with diabetic kidney injury. However, S100A9 expression was down-regulated in kidney tissue after treatment with Shenjiankang, which indicated that Shenjiankang had a good regulation of S100A9 gene expression Role, its mechanism remains to be further studied.