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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的作用机制,筛选药效作用靶点,为临床推广应用提供科学依据。[方法]本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、逆转录聚合酶链反应法(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]鸟嘌呤核苷酸结合蛋白α12(Gna12),在糖尿病大鼠肾脏表达下调,肾消康中剂量组表达上调。[结论]应用AffymetrixRat2302.0基因表达谱芯片检测糖尿病大鼠肾脏基因的表达,Gna12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。基因功能分析表明,在糖尿病状态下,Gna12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Gna12表达上调,可见肾消康对Gna12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 curative effect of traditional Chinese medicine Shenji Decoction on diabetic nephropathy rats and the effect on kidney gene expression, explore the pathogenesis of diabetic nephropathy (DN), reveal the mechanism of traditional Chinese medicine Shenji Decoction in preventing and treating DN, and screen the efficacy The role of target for clinical application to provide a scientific basis. [Method] In this experiment, streptozotocin (STZ) plus high-calorie diet was used to establish the model of kidney injury in diabetic rats. By using radioimmunoassay, biochemistry, light microscopy, electron microscopy and Affymetrix gene expression microarray and reverse transcription-polymerase chain reaction (RT-PCR) and other advanced detection methods and methods, observed a number of indicators, especially by cluster analysis Looking for genes with important relevance to DN and further using real-time fluorescence quantitative RT-PCR to do validation experiments. [Results] Guanine nucleotide binding protein α12 (Gna12) was down-regulated in the kidney of diabetic rats and up-regulated in the middle-dose Shenzhikang group. [Conclusion] AffymetrixRat2302.0 gene expression microarray was used to detect the expression of renal genes in diabetic rats. Gna12 was one of the differentially expressed and pharmacologically active targets of kidney, and the gene was analyzed by real-time RT-PCR. Gene function analysis showed that in the state of diabetes, the expression of Gna12 was down-regulated, and it had an important correlation with diabetic nephropathy. The expression of Gna12 was up-regulated in kidney tissue after treatment with Shenkang Qikang, which indicated that Shenjiankang had a good regulation of Gna12 gene expression Role, its mechanism remains to be further studied.