五氯硝基苯影响大鼠卵泡发育的机制研究  

Mechanism research of effect of pentachloronitrobenzene on follicular development in rats

作　　者：淡清华 骆海燕 张雨彤 高小博 陆彩玲 DAN Qing-hua;LUO Hai-yan;ZHANG Yu-tong;GAO Xiao-bo;LU Cai-ling(Department of Genetics,National Research Institute for Family Planning,Beijing 100081;Graduate School,Peking Union Medical College,Beijing 100005)

机构地区：[1]国家卫生健康委科学技术研究所遗传优生中心,北京100081 [2]北京协和医学院研究生院,北京100005

出　　处：《生殖医学杂志》2022年第4期506-513,共8页Journal of Reproductive Medicine

基　　金：国家自然科学基金项目(81970343,82070327);中央级公益性科研院所基本科研业务项目(2020GJZ09,2021GJZ04)。

摘　　要：目的探究有机氯农药五氯硝基苯(PCNB)影响大鼠卵泡发育的分子机制。方法将14只青春前期SD雌性大鼠随机分为对照组和实验组,每组各7只。实验组给予PCNB(100 mg·kg^(-1)·d^(-1)),对照组给予等体积玉米油,连续灌胃7 d。取卵巢组织HE染色,观察并统计各级卵泡数目;通过RNA-Seq测序筛选差异表达基因,随后进行GO富集和KEGG功能注释分析;Western Blot检测大鼠卵巢AKT的磷酸化水平和张力蛋白同源物(PTEN)的表达。结果与对照组相比,实验组原始卵泡显著减少(P<0.05),生长卵泡显著增加(P<0.05)。测序结果共筛选出261个差异表达的基因(|log2 Fold change|>1,q<0.05),与对照组相比,实验组上调的基因有62个,下调的基因有199个;GO分析显示差异表达基因主要富集在细胞分化、细胞内信号转导、ATP结合的微管装配等;KEGG富集分析提示PI3K-AKT信号通路富集的差异基因数目较多;Western Blot结果显示,与对照组相比,实验组AKT磷酸化水平显著增加(P<0.05);PTEN表达量显著减少(P<0.05)。结论PCNB加速大鼠原始卵泡发育与卵巢PI3K-AKT信号通路激活相关。Objective:To investigate the molecular mechanism of the effects of organic chlorine pesticide pentachloronitrobenzene(PCNB)on follicle development in rats.Methods:Fourteen prepubertal female SD rats were randomly divided into control group and experimental group with 7 rats in each group.The rats of experimental group were given PCNB(100 mg·kg^(-1)·d^(-1)),and the rats of control group were given equal volume of corn oil,for continuous gavage 7 days.The number of follicles at all levels was counted and observed by HE staining.Differentially expressed genes were screened by RNA-Seq sequencing,followed by GO enrichment and KEGG functional annotation analysis.The phosphorylation of AKT and the expression of PTEN were detected by Western Blot.Results:Compared with the control group,the primordial follicles in the experimental group were significantly decreased(P<0.05),but the growth follicles were significantly increased(P<0.05).RNA-Seq sequencing screened 261 differentially expressed genes(|log2 fold change|>1,q<0.05).Compared with the control group,62 up-regulated genes and 199 down-regulated genes were found in the experimental group.GO enrichment analysis showed that differentially expressed genes were mainly concentrated in cell differentiation,intracellular signal transduction and ATP-bound microtubule assembly.KEGG enrichment analysis indicated that there were more differential genes enriched in PI3K-AKT signaling pathway.Western blot showed that the phosphorylation level of AKT was significantly increased compared with the control group(P<0.05),but expression of PTEN protein was significantly decreased(P<0.05).Conclusions:PCNB accelerates the development of primordial follicles in rats,which was related to the activation of PI3K-AKT signaling pathway in rats ovary.

关 键 词：五氯硝基苯 卵巢 卵泡发育 PI3K-AKT 

分 类 号：R994.6[医药卫生—毒理学]