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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 effect and mechanism of genistein on the differential expression of ovarian epithelial cells in premature ovarian failure (POF) induced by CDDP. Methods According to the literature, the mouse model of hepatic-kidney yin deficiency syndrome with low ovarian function after chemotherapy was established. The total RNA of POF group and normal control group’s ovarian tissue was extracted by one-step method. Cy3 and Cy5 were used for reverse transcription. Fluorescent labeling was performed to obtain probes for mouse ovarian cDNAs in each group and then hybridized with cDNA gene expression profiling chips. The results were scanned by a laser scanner and analyzed by software for image analysis, normalization, ratio analysis, and cluster analysis. RESULTS: Gene expression profiling analysis after GEN administration revealed 60 differentially expressed genes, of which 22 genes were increased and 38 genes were decreased. Functional classification of differentially expressed genes mainly focuses on three major categories of cellular components, molecular functions, and biological processes. Pathway analysis results showed that GEN is mainly involved in the differential expression process of large ribosome and small ribosome, and has a significant effect on the regulation of gene network expression of GnRH and MAPK pathways. Conclusion In this study, a large number of differentially expressed genes were screened from ovary tissue of POF mice after GEN administration. GEN may partially relieve the restriction of RNA function in mice with liver-kidney yin deficiency syndrome, increase the expression of ATP metabolism-related genes, and enable energy metabolism. The effect is enhanced to improve ovarian function.