肾消康对糖尿病肾病大鼠肾组织基因S100A9表达的影响

Effect of ShenXiaoKang on Gene-S100A9 Expression in Diabetic Rats

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 therapeutic effect of a complex prescription of Shenxiaokang on DN rats and its mechanism. Methods The model of DN was 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, Af- fymetrix Co. US. Rat 2302.0 gene expression profiling chip ,and real time quantitative reverse transcription polymerase chain reac- tion technique to observe the effect of Shenxiaokang on indicators. Results S100A9 gene was in the DN model group rat kidneys and down - regulated in the Shenxiaokang treatment group rat kidneys. Conclusion S100A9 gene was one of the renal tissue difference expresstion and drug effect targets and it was analyzed by real time quantitative reverse transcription polymerase chain reaction technique. It was the first time at home and abroad. The expression of SI00A9 gene is associated with diabetic kidney damage, and Shenxiaokang has benign effect on S100A9 gene Expresstion.