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目的观察中药复方肾消康对糖尿病肾病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(DN)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。结果真核生物翻译起始因子3,8亚基(eIF3s8),在糖尿病大鼠肾脏表达下调,肾消康治疗组表达上调。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,eIF3s8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,eIF3s8表达下调,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eIF3s8表达上调,可见肾消康对eIF3s8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 rats and its effect on gene expression of kidney, and to explore the pathogenesis of diabetic nephropathy (DN), to reveal the mechanism of traditional Chinese medicine Shenji Decoction in preventing and treating DN, and to screen the pharmacodynamic target Point, to provide a scientific basis for clinical application. Methods Streptozotocin (STZ) plus high-calorie diet was used to establish a model of kidney injury in diabetic rats. 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 eukaryotic translation initiation factor 3,8 subunit (eIF3s8) was down-regulated in the kidney of diabetic rats and was up-regulated in the Shenkukang treatment group. Conclusions Affymetrix Rat2302.0 gene expression microarray was used to detect the expression of renal genes in diabetic rats. EIF3s8 was one of the differentially expressed and pharmacologically active targets of kidney, and the gene was analyzed by RT-PCR. This is the first time at home and abroad. Gene function analysis showed that in the diabetic state, eIF3s8 expression down regulation, and diabetic kidney injury have important correlation, and after the treatment of renal decongestant rat eIF3s8 upregulation of kidney tissue, can be seen Shenjiankang eIF3s8 gene expression has a benign regulation Role, its mechanism remains to be further studied.