基因芯片筛选抗胸腺细胞血清性肾炎大鼠肾组织基因表达差异的初步研究

Exploration of gene status in renal tissues of rats with anti-thymocyte serum nephritis by genechip

目的:利用基因芯片检查大鼠系膜增生性肾炎,即抗胸腺细胞血清性肾炎(ATSN)模型大鼠发病40 m in和5 d时肾组织相关基因的表达情况,探讨ATSN大鼠不同时相肾组织基因表达的差异并作功能分析。方法:用抗胸腺细胞抗血清(ATS)复制大鼠ATSN模型,抽提发病40 m in和5 d时肾组织RNA,分别用逆转录合成荧光分子掺入的cDNA作探针进行基因芯片杂交。用Agilent扫描仪扫描,Im agene软件读取数据,最后用Genespring进行norm alize处理和差异表达基因的筛选。差异基因筛选标准为实验组/对照组比值(ratio)>或=2为上调基因,实验组/对照组ratio<或=0.5为下调基因。然后选取上调和下调基因登录GenBank查找其相关功能。结果:在9 234个大鼠基因中(去除质控基因),ATSN大鼠40 m in时肾组织上调基因为341个,其中部分为凋亡相关的调控基因,部分为增生相关基因及一些与信号转导有关的基因;下调基因为392个,其中一些是酶类基因,部分为生长因子和细胞因子受体基因等,其余大多上调和下调基因功能不详。ATSN发病5 d时,肾组织上调基因是213个,包括增生相关基因、信号转导相关基因等;下调基因119个,其中部分为酶类和细胞因子相关基因。5 d时上调和下调基因大多功能不清楚。结论:大鼠ATSN发病早期(40 m in),致病相关基因已被激活(如凋亡和增生相关基因),其中相关信号转导分子基因也表达上调,而在ATSN大鼠发病5 d时,基因表达谱有所改变,其中涉及的基因多为增生相关基因和信号转导相关基因。

AIM： To explore gene expression in renal tissues from rats with mesangial proliferative glomerulonephritis （MPGN） , namely, anti- thymocyte serum nephritis （ATSN） , at 40 minutes and 5 days after injection with rabbit anti - thymocyte serum （ATS） , and to analyze the differences of gene spectrum and functions between the two phases of ATSN with genechip. METHODS ： The rat model of ATSN was reproduced by injection of ATS to the animals. 40 minutes and 5 days later, the total RNA in renal tissues was extracted. Further, genechip probe produced by reverse transcription with fluorescence mixed into the first chain of cDNA from the two phases RNA was hybridized with the rat cDNA expression sequence chip severally. The chips were subsequently scanned by Agilent scanner and read with Imagene software. In succession, normalized data and the ratio of the gene fluorescent intensity in ATSN and the control groups were performed with Genespring, by which the down -regulated genes with ratios equal or lower 0. 5 and the up -regulated genes with ratios equal or higher than 2 were screened out. Finally, the selected up - regulated and down - regulated genes were searched in GenBank to find the relative functions. RESULTS： There were 341 increased genes, including apoptosis, proliferation and signal transduction related genes at 40 minutes after ATS injection. 392 decreased genes containing a number of enzyme related genes, growth factors and cytokine receptors, etc, were also observed. In addition, there were 213 increased genes and 119 lessened genes at 5 days of ATSN, while most of the functions of many genes remained unclear. CONCLUSIONS： At the early period of the ATSN （40 min）, some nosogenetic genes, such as apoptotic and proliferative genes as well as several signal transduction - related genes are activated and expressed significantly, while the profile of gene expression at 5 days, such as many proliferative genes or transduction - related genes, are changed.