摘要
目的 探讨Perilipin基因差异表达与雄激素缺乏之间的关系.方法 采用D-半乳糖注射的方法建立大鼠亚急性衰老雄激素缺乏动物模型,检测血清总睾酮（TT）、超氧化物岐化酶（SOD）、丙二醛（MDA）水平判断造模成功,采用Percoll非连续密度梯度离心法分离睾丸间质细胞（Leydig）,原代培养72h后提取大鼠Leydig细胞总RNA,基因芯片筛选差异表达基因,最后半定量RT-PCR验证芯片结果.结果 与对照组比较,模型组（低剂量组和高剂量组）血清TT浓度、SOD活力下降,MDA含量升高,差异有统计学意义；基因芯片结果显示,脂类代谢途径与雄激素缺乏最为相关,差异表达基因分别为Perilipin,Fabp4,Mgll;半定量RT-PCR结果发现,Perilipin基因及其两个亚型表达显著下调.结论 在RNA水平Perilipin基因表达与雄激素缺乏可能有关.
Objective To evaluate the relationship of Perilipin expression with reduced androgen synthesis in aging male rats. Methods The three-month old rats were injected every day with D-galactose （300 mg/kg） for two months to establish the model of aging rat. The levels of total testosterone （TT）, superoxitase dismutase （SOD） and malondialdehyde （MDA） in serum were used as the criterion to evaluate the quality of the animal model. For measurement of differentially-expressed genes, the Leydig cells was separated by Percoll discontinuous density gradient centrifugation, cultured for three day and then used for preparation of total RNA. Gene expression profile in the aging Leydig cells was studied in by cDNA microarray technique plus validation of screened genes by RT-PCR. Results In D-galactose- injected rats, the serum levels of MDA and SOD were significantly higher than those of the control rats. However, the serum level of TT was significantly lower in the rats injected with D-galactose than that of the control rats. Expression of hundreds of genes changed in the Leydig cells of aging rats, compared to that in the control, screened by cDNA microarray technique. Among these genes, Perilipin expression was selected for validation by RT-PCR technique, and our data indicated that levels of both Perilipin A and Perilipin B expression were significantly reduced in the aging Leydig cells. Conclusion The aging male rats were well established. Down-regulated expression of Perilipin may result in a reduced androgen synthesis in Leydig cells of aging male rats.
出处
《实验动物与比较医学》
CAS
2011年第6期405-409,共5页
Laboratory Animal and Comparative Medicine
基金
上海市科委科研项目（09DJ1400400,09140903200）
