目的利用基因芯片技术研究染料木黄酮(Genistein,Gen)对体外培养的小鼠睾丸组织全基因表达谱的影响,从基因水平分析Gen影响睾丸细胞增殖凋亡的作用机制。方法利用旋转通气法体外培养乳鼠睾丸组织,给予5μmol/LGen干预,同时设立DMs0溶剂对照组。培养72h后提取组织总RNA,用单标Agilent小鼠全基因芯片检测两组睾丸组织基因表达,筛选差异基因,进行基因功能分类。结果与对照组比较,Gen组筛选出84个表达差异有统计学意义的基因(P〈0.05),其中47个基因表达上调,37个基因表达下调。按差异基因功能富集分析(G0)分类标准进行分类,筛选出有统计学意义的GO分类14个(P〈O.05),主要涉及生殖发育、细胞增殖、细胞周期等功能。结论通过全基因芯片检测发现,Gen主要影响睾丸组织生殖发育、细胞增殖和细胞周期等功能相关基因表达,从而影响精子发生和睾丸细胞的增殖和凋亡,影响睾丸的功能。
Objective To investigate the effects of Genistein (Gen) on the whole expression profiles of testes in mice by using the gene chip technology, and analyze the mechanism how Gen exerts testis on gene level. Methods The testicular tissues of suckling mice were cultured in vitro, and separated into control group and Gen group with the dose of 5 p.mol/L. After 72 h of culture, we extracted the total RNA and purified it, then detected the gene expression by using Agilent gene array. Results Compared with the control, there were 84 genes expressed differently in Gen group, including 47 up-regulated genes and 37 down-regulated genes. We classified these genes as 14 categories in Gen group by GO (P〈0.05), including cell proliferation, cell death, the immune system and reproductive development. Conclusion The whole gene chip test found that Gen can mainly affect the functions of testes by regulating the expression of gene related to cell development, proliferation and cell cycle function, which can influence the spermatogenesis and change the testis cell proliferation and apoptosis.