肾脏基因表达数据库的建立与肾脏衰老相关基因表达谱研究

Gene expression profile of aging and construction of a kidney expression database

目的 运用基因芯片技术及公共数据库，首次建立大鼠肾脏基因表达谱数据库。比较成年及老年大鼠肾脏基因表达变化，筛选肾脏衰老相关基因。方法 将代表８ １９２条基因的ＰＣＲ产物用 Ｃａｒｔｅｓｉａｎ Ｐｉｘｓｙｓ ７５００点样仪按微矩阵排列点样于 Ｄａｋｏ玻璃片，将等量的成年大鼠肾脏组织和衰老大鼠肾组织ｍＲＮＡ分别用ｃｙ－３、ｃｙ－５逆转录荧光标记，制作ｃＤＮＡ探针，混合后与上述表达谱芯片进行杂交。Ｓｃａｎ Ａｒｒａｙ ３０００扫描仪扫描杂交信号荧光强度，ＩｍａＧＥＮＥ３．０软件分析扫描结果。与公共数据库比较，建立大鼠肾脏表达谱数据库。结果 在进入研究的８ １９２条基因中，１２４条基因（１．５％）在衰老大鼠肾脏组织中有表达差异（１．７倍以上）。在老年大鼠肾组织中表达增加的基因有７１条，其中１６条（２２．５％）为应激反应相关或炎症相关基因，１７条（２３．９％）为细胞外基质代谢／细胞骨架／肿瘤相关基因。在衰老大鼠肾组织表达下降的基因有５３条，其中９条（１６．９％）为能量代谢相关基因，１２条（２２．６％）为蛋白质合成／代谢相关基因，６条（１１．３％）为细胞周期／有丝分裂相关基因，４条（７．５％）为ＤＮＡ／ＲＮＡ损伤修复基因。

Objective To construct a gene expression profile database and to screen the candidate aging-related genes by using microarray analysis. Methods PCR products on behalf of 8 192 genes from mixed tissue library were assembled on the modified Dako glass slide. The mRNA from adult and aging rats were labelled wiht cy-3 and cy-5 separately by RT-PCR as cDNA probes. Then two probes were mixed and hybridized with the above-mentioned gene chip. Scan array 3000 was used for scanning the hybridizing signals and ImageGENE 3. 0 for data analysis. Results Of the 8 192 genes monitored, 124 genes (1. 5% ) showed consistent changes in expression level of at least 1. 7 fold. The expression of 71 genes (57. 2% ) was up-regulated and 53 genes(42. 8% ) was down-regulated. Conclusions These results provide insight into the mechanism of aging. The characteristic appearance of aging-related genes expression includes increased level of stress response and inflammation, increased level of ECM/cytoskeleton decreased DNA/RNA repairment and decreased protein assembling.