维生素D缺乏模型小鼠子宫转录组的测序分析  

Transcriptome sequencing of the uterine in a mouse model of vitamin D deficiency

作　　者：陈霞[1] 商宇伟 王林晓[2] 史轶超[1] 刘慧君 孙慧婷[1] Chen Xia;Shang Yuwei;Wang Linxiao;Shi Yichao;Liu Huijun;Sun Huiting(Department of Reproductive Center,Changzhou Second People’s Hospital Affiliated to Nanjing Medical University,Changzhou 213003,Jiangsu Province,China;Laboratory of Neurological Diseases,Changzhou Second People’s Hospital Affiliated to Nanjing Medical University,Changzhou 213003,Jiangsu Province,China)

机构地区：[1]南京医科大学附属常州市第二人民医院生殖中心,江苏省常州市213003 [2]南京医科大学附属常州市第二人民医院神经疾病研究室,江苏省常州市213003

出　　处：《中国组织工程研究》2022年第26期4101-4106,共6页Chinese Journal of Tissue Engineering Research

基　　金：常州市科技局应用基础研究计划(CJ20200103),项目负责人:孙慧婷;常州市科技计划资助项目(CJ20210110),项目负责人:陈霞;国家自然科学基金青年基金项目(81803498),项目负责人:王林晓。

摘　　要：背景:近来发现维生素D受体广泛分布在卵巢、子宫等女性生殖系统,与自然流产及妊娠期糖尿病等不良妊娠结局密切相关,但其具体机制尚不清楚。目的:通过对维生素D缺乏小鼠子宫组织进行全转录组分析,寻找潜在差异表达miRNA及相关调控网络。方法:取雌性C57BL/6J小鼠18只随机分为2组,维生素D缺乏组给予维生素D缺乏饲料喂养,正常对照组给予维生素D充足饲料喂养,每周记录小鼠体质量。8周后检测小鼠血清中25,(OH)D3和激素水平,对小鼠子宫组织进行苏木精-伊红染色并收集两组子宫组织进行转录组测序,进行基因本体、京都基因与基因组百科全书生物信息学分析。结果与结论:①两组小鼠间体质量无明显差异;与正常对照组相比,维生素D缺乏组小鼠血清25,(OH)D3、雌二醇水平显著降低,睾酮水平显著升高;②子宫组织苏木精-伊红染色结果显示,维生素D缺乏组子宫内膜皱褶减少;③转录组测序结果共筛选出差异表达的miRNA 25个,其中上调9个(如miR-541-5p和miR-205-5p),下调16个(如miR-378d和miR-708-5p);④同时维生素D缺乏小鼠在多项基因本体富集类别中具有显著差异,如发育过程、细胞成分组织或生物发生等;此外,京都基因与基因组百科全书功能富集结果显示,差异表达基因多与生化代谢途径和信号转导途径相关,且些富集通路中的差异表达基因主要影响了PI3K-Akt信号通路、MAPK信号通路、胰岛素信号通路、促性腺激素释放激素信号通路及卵母细胞减数分裂通路,与生殖系统调控密切相关;⑤结果表明,维生素D缺乏可导致雌鼠子宫着床条件变差,并与调节小鼠生殖系统胰岛素等多个信号通路相关。BACKGROUND:It has been recently discovered that vitamin D receptor is widely distributed in the female reproductive system such as the ovary and uterus,which is closely related to adverse pregnancy outcomes such as spontaneous abortion and gestational diabetes mellitus,but its specific mechanism is still unclear.OBJECTIVE:To investigate potential differentially expressed miRNAs and related regulatory networks via a whole transcriptome analysis of the uterine tissue of vitamin D-deficient mice.METHODS:Eighteen female C57BL/6J mice were randomly divided into two groups:a vitamin D deficiency group and a normal control group.Mice in the vitamin D deficiency group were fed with vitamin D deficiency diet,and mice in the normal control group were fed with vitamin D sufficiency diet.The body mass of the mice was recorded every week.After 8 weeks,the serum levels of 25,(OH)D3 and hormones in the mice were tested.Hematoxylin-eosin staining was used to detect mouse uterine tissue.Uterine tissue samples of two groups were collected for transcriptome sequencing,and bioinformatics analyses of gene ontology and Kyoto encyclopedia of genes and genomes.RESULTS AND CONCLUSION:There was no difference in body mass between the two groups.Compared with the normal control group,the levels of 25,(OH)D3 and estradiol in mouse serum were significantly reduced,and the testosterone level was significantly increased in the vitamin D deficiency group.Hematoxylin-eosin staining of uterine tissue showed a reduction in the endometrial folds of mice in the vitamin D deficiency group.Twenty-five differentially expressed miRNAs were screened out by transcriptome sequencing,of which 9 were up-regulated(such as miR-541-5p and miR-205-5p)and 16 were down-regulated(such as miR-378d and miR-708-5p).At the same time,vitamin D deficient mice showed significantly differences in multiple gene ontology enrichment categories,such as developmental process,cell composition,tissue or biogenesis.In addition,the functional enrichment results of Kyoto genes and ge

关 键 词：维生素D缺乏 子宫 转录组测序 miRNA 生殖系统 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]