肾消康对糖尿病大鼠肾组织基因Gna12表达的影响

Effect of Shenxiaokang on the renal tissues expression of Gene Gna12 in diabetic nephropathy rats

关键词:  肾消康  糖尿病  肾脏损伤  基因表达谱  鸟嘌呤核苷酸结合蛋白α12

Key words:  Shenxiaokang  diabetes  diabetic kidney damage  gene expression profiling  Guanine nucleotide binding protein alpha 12

摘要:

[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Affymetrix Rat2302.0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。

Abstract:

[Objective] To observe the therapeutical effect of a complex prescription of Shenxiaokang on DN rats and discuss the mechanism of action in the treatment of DN, then provide experimental data for clinical practice. [Methods] The model rats male Wistar were established with multiple injections of low dose STZ and high calorie diet. Kinds of examination methods were applied, including radiommunoassay, biochemistry, light microscope, electron microscrope, immunohistochemical, Affymetrix Co.US. rat 2302.0 gene expression profiling chip, and real time quantitative reverse transcription polymerase chain reaction technique to observe the effect of Shenxiaokang on indicators. Most especially applied cluster analysis method to make certain important associated genes with DN, and further applied RT-PCR to do confirmatory experiment. [Results] Gna12 gene was down-regulation in the DN model group rat kidneys and up-regulation in the Shenxiaokang treatment group rat kidneys. [Conclusion] Applied Affymetrix Co.US. rat 2302.0 gene expression profiling chip to test diabetic nephropathy rats renal tissues genes expresstion, Gna12 gene is one of the renal tissues difference expresstion and drug effect targets and applied real time quantitative reverse transcription polymerase chain reaction technique to analyze it. It is the first time at home and abroad. To genes function analyze show that, the Gna12 gene is down-regulation in the DN model group rat kidneys. It is important associated with diabetic kidney damage, and up-regulation in the Shenxiaokang treatment group rat kidneys. Shenxiaokang has benign effect on Gna12 gene Expresstion. Its mechanism remains to be studied further.