金雀异黄素对肝肾阴虚证卵巢功能早衰小鼠基因差异表达作用研究

Effects of genistein on ovary gene expression pattern of CDDP induced premature ovarian failure mice with syndrome of kidney and liver yin asthenia

目的探讨金雀异黄素(GEN)对CDDP致肝肾阴虚证卵巢功能早衰(premature ovarian fail-ure,POF)小鼠卵巢基因差异表达的影响及机制。方法依照文献建立化疗损伤后卵巢功能低下肝肾阴虚证小鼠模型,按一步法抽提GEN给药后POF组和正常对照组小鼠卵巢组织的总RNA;经逆转录分别用Cy3、Cy5荧光标记,获得各组小鼠卵巢cDNA的探针,再与cDNA基因表达谱芯片杂交,结果由激光扫描仪扫描并用软件进行图像分析、标准化处理、ratio值分析、聚类分析。结果GEN给药后基因表达谱分析发现60个差异表达基因,其中22个基因表达增加,38个基因表达下降。差异表达基因功能分类主要集中于细胞组件、分子功能和生物学过程三大类。pathway分析结果表明,GEN主要参与大核蛋白体和小核蛋白体差异表达过程,对GnRH、MAPK等通路的基因网络表达调控作用明显。结论本研究从GEN给药后的POF小鼠卵巢组织中筛选出大量的差异表达基因,GEN可能部分解除肝肾阴虚证小鼠RNA功能受限,提升ATP代谢相关基因表达作用,使能量代谢作用得到增强,进而改善卵巢功能。

Objective To investigate the effects of genistein（GEN） on ovary genes differential expression in CDDP induced premature ovarian failure（POF） mice with syndrome of kidney and liver yin asthenia,and its mechanism.Methods POF mouse model with syndrome of kidney and liver yin asthenia was established by chemotherapy injury according to literature.The ovary total-RNAs of GEN group and normal control group were extracted and were reversely transcribed with Cy3,Cy5 fluorescent labeling to obtain cDNA probes that were hybridized with cDNA microarray chips.The results from the laser scanner were made image analysis,standardization,ratio analysis and cluster analysis.Results Sixty genes expressed were screened out differentially;22 were up-regulated and 38 were down-regulated.These Genes were classified into three groups,which were cell composition,molecule function and biology process.Pathway analysis indicated that GEN participated in the process of ribosome gene expression and controlled the GnRH and MAPK gene expression mainly.Conclusiosn There are many different gene expression pattern in POF mouse with syndrome of kidney and liver yin asthenia.GEN may remove functional restriction of RNA partly and up-regulated ATP metabolism related gene expression,which will strengthen energy metabolism.