多精受精中人卵丘颗粒细胞差异基因表达分析  

Analysis of differential gene expression of cumulus granulosa cells in polyspermy

作　　者：王丽[1] 赵杰[1] 陈秀娟[1] 杜琛[1] 刘芳[1] 何江峰 梅步俊[3] WANG Li;ZHAO Jie;CHEN Xiu-juan;DU Chen;LIU Fang;HE Jiang-feng;MEI Bu-jun(The Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010050;Inner Mongolia Academy of Agriculture&Animal Husbandry Science,Hohhot 010030;Hetao College,Bayannur 015000)

机构地区：[1]内蒙古医科大学附属医院,呼和浩特010050 [2]内蒙古自治区农牧业科学院,呼和浩特010030 [3]河套学院,巴彦淖尔015000

出　　处：《生殖医学杂志》2022年第11期1554-1560,共7页Journal of Reproductive Medicine

基　　金：内蒙古医科大学附属医院科技成果转化专项(2022NYFYCGZH001);内蒙古自治区科技计划项目(2021GG0030);国家自然科学基金(31760660);内蒙古自治区自然科学基金项目(2019MS08121,2020LH03017);内蒙古教育厅研究生教育教学改革研究与实践项目(YJ20191013206)。

摘　　要：目的 筛选人卵丘颗粒细胞中影响卵母细胞多精受精的差异表达基因。方法 筛选3名因输卵管因素导致继发不孕的患者为研究对象,长方案促排卵后,在授精前10 min,对每个卵母细胞外围的卵丘颗粒细胞进行切割,分装无菌管保存。卵冠丘复合体(OCCC)进行微滴授精,受精后17~18 h,观察受精情况;并根据卵母细胞受精情况分为多精受精组和正常受精组,收集两组对应的颗粒细胞。提取颗粒细胞总mRNA,构建cDNA文库并进行测序,将得到的数据进行生物信息学分析,筛选出差异表达基因,并将差异表达基因进行基因本体论(GO)分析和京都基因和基因组数据库(KEGG)分析。结果 与正常受精组比较,多精受精组中显著上调的基因有106个,显著下调的基因有33个。上调的基因主要涉及P53信号通路、PI3K-AKT信号通路、肌动蛋白细胞骨架信号通路和HIPPO信号通路,其中基因BMP2、SESN3、CDK6、GRLF1(ARHGAP35)与多精受精有关。结论 与正常受精比较,多精受精时颗粒细胞存在差异表达的基因,对这些基因的进一步了解可能为后续开展异常受精的影响因素分析提供一定参考。Objective:To screen differentially expressed genes which affected polyspermy in human cumulus granulosa.Methods:Three patients with secondary infertility caused by fallopian tube factor were chosen as study objects.After ovulation stimulate with long protocol,the cumulus granulosa cells in the peripheral part of each oocyte were obtained 10 min.before fertilization and divided into sterile tubes for preservation.Oocyte-corona-cumulus complex(OCCC)was fertilized by microdroplet,fertilization changes were observed at 17 to 18 hours after fertilization.According to the fertilization status,the oocytes were divided into polyspermy group and normal fertilization group.The corresponding granular cells of the two groups were collected,and total mRNA was extracted from cumulus granulosa cells for cDNA library construction.The cDNAs were sequenced and the obtained data were analyzed by bioinformatics to screen out differentially expressed genes.Differential expression genes were analyzed by gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG).Results:Compared with normal fertilization group,106 genes were significantly up-regulated and 33 genes were significantly down-regulated in polyspermy group.The up-regulated genes mainly involved in P53 signaling pathway,PI3K-AKT signaling pathway,actin cytoskeleton signaling pathway and HIPPO signaling pathway.Among them,BMP2,SESN3,CDK6,GRLF1(ARHGAP35)may be associated with polyspermy.Conclusions:There are differentially expressed genes in granulosa cells during polyspermy.Further understanding of these genes may provide some reference for the subsequent analysis of factors affecting abnormal fertilization.

关 键 词：卵丘颗粒细胞 卵母细胞 受精 差异基因表达 

分 类 号：R711.6[医药卫生—妇产科学]