目的:比较糖尿病肾病（DN）大鼠和中药复方肾消康治疗组大鼠肾脏组织中基因表达的差异,并用Eve green实时荧光RT-PCR技术定量检测CcL5mRNA在各组大鼠肾脏组织的表达水平。方法:（1）差异表达基因的筛选:实验大鼠分为空白组、模型组、治疗组。从肾脏组织中抽提mRNA,由纯化的总RNA合成双链cDNA,并将其纯化,以生物素标记cRNA合成,纯化和质控体外转录（IVT）产物,片断化cRNA,然后进行杂交、洗脱、染色、芯片扫描并用软件进行统计分析,筛选出差异表达数据。不同肾脏组织样本用灰度扫描图和杂交信号强度散点图表示。（2）荧光定量RT-PCR的检测: RNA样品反转录,荧光定量PCR反应。结果:仅对筛选出差异表达基因谱中的CcL5mRNA进行报告。糖尿病肾病模型组肾脏组织基因CcL5mRNA表达上调,中药复方肾消康治疗组肾脏组织基因CcL5mRNA表达下调。结论:CcL5mRNA在糖尿病状态下肾脏也有表达,并且表达上调,而中药复方肾消康可使其下调,这也是中药复方对糖尿病性肾组织损伤具有保护作用的机制之一。

Objective: studying genetic expression differences in renal tissues between the rats having diabetic nephropathy and the rats treated with the medical herbs. CcL5mRNA- expression in renal tissues is measured, using Evergeen real time RT-PCR. Method: 1) screening out the genes that are expressed differently: rats are divided into three groups for comparison. Extracting mRNA from renal tissues, and making them into duplex cDNA, using purified RNA. Labeling cRNA with biotin, sublimating transcript products in vitro, and fragmenting cRNA. Hybridizing, washing, staining and scanning are also used to complete the process. A computer program is used to sort out expression differences. Renal tissue samples are labeled using either grey degree scanning, or a scattering graph. 2) Measuring RNA reverse using Evergeen real time RT-PCR. Result: this paper only reports differed CcL5 mRNA expressions. A raised CcL5mRNA expression can be seen in the genes of untreated diabetic rats, while the same expression is reduced in the treated one. Conclusion: CcL5mRNA finds a raised expression in untreated diabetic rats. Fortunately, ShenXiaoKang cuts down such expressions, which exhibits a merit that medical herbs can bring to protecting renal tissues from further damage.

黑龙江杰出青年科学基金项目(2070222):肾消康对糖尿病肾损伤保护作用的基因组学研究,负责人:姜德友。