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 therapeutic 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 male model Wistar rats 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.Cluster analysis method especially applied was to make certain important associated genes with DN,and further applied RT-PCR to do confirmatory experiment. Results eIF3s8 gene was down-regulated in the DN model group and up-regulated in the Shenxiaokang treatment group. Conclusion It was the first time at home and abroad to apply Affymetrix Co.US.Rat 2302.0 gene expression profiling chip to test diabetic nephropathy rats renal tissues genes expresstion in which.eIF3s8 gene was one of the renal tissues difference expresstion and drug effect targets,and to apply real time quantitative reverse transcription polymerase chain reaction technique to analyze it.Gene function analysis show that,the eIF3s8 gene is down-regulated in the DN model group rat kidneys.It is important associated with diabetic kidney damage,and up-regulated in the Shenxiaokang treatment group rat kidneys,Shenxiaokang has benign effect on eIF3s8 gene expresstion.Its mechanism remains to be studied further.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2012年第1期62-64,共3页
Lishizhen Medicine and Materia Medica Research
基金
黑龙江省杰出青年自然科学基金(No.207022)
关键词
肾消康
糖尿病
肾脏损伤
基因表达谱
真核生物翻译起始因子3
8亚基
Shenxiaokang Diabetes Diabetic kidney damage Gene expression profiling Eukaryotic translation initiation factor 3 subunit 8
