容受前期与容受期子宫内膜基因表达及代谢物水平差异分析

Analysis of the difference in metabolites and gene expressions between pre-receptive and receptive endometria

目的分析容受前期和容受期子宫内膜基因表达及宫颈分泌物中氨基酸和肉碱水平差异,为探讨新的子宫内膜容受性分子标记物提供线索和依据。方法选取2020年1月6号至2022年1月31日就诊于临沂市人民医院生殖医学科的共59例不孕女性患者作为研究对象,收集临床资料。匹配年龄、体质量指数及不孕年限等因素选取准备胚胎移植的不孕女性3对(6名),取内膜组织进行基因转录表达分析。另选经辅助生殖技术妊娠25例为对照组,门诊监测排卵未妊娠女性28例为病例组,采用超声判定内膜容受性状态。前者提取子宫内膜组织进行测序后,进行差异表达基因的GO数据库和KEGG数据库富集分析。后者取对照组和病例组宫颈分泌物,经质谱检测其中氨基酸和肉碱水平。采用秩和检验、t检验及χ^(2)检验进行统计学分析。结果研究对象基本临床资料无明显差异(P>0.05)。基因测序提示上/下调最显著的基因分别是HLA-DRB5和MMP10。GO和KEGG富集差异表达基因较多的生物学过程、分子功能和通路主要有免疫调控、细胞粘附和色氨酸代谢等。分泌物代谢分析发现多种氨基酸和肉碱代谢物水平在二组间差异明显(P<0.05),主要为氨基酸和肉碱水平如丙氨酸、缬氨酸、3-羟基丁酰肉碱(C4OH)+丙二酰肉碱(C3DC)/葵酰肉碱(C10)等。结论容受前期与容受期子宫内膜组织相比,基因转录表达差异显著,可能与子宫内膜容受性状态存在关联。氨基酸和肉碱水平提示差异变化较大指标可能作为标记物预测子宫内膜容受性状态。

Objective To analyze the difference in the gene expression,amino acid and carnitine levels in the cervical secretions between the endometria of pre-receptive and receptive stages,with an aim to provide clues for identifying new molecular markers for endometrial receptivity.Methods Fifty nine infertile women treated at the Department of Reproductive Medicine of Linyi People′s Hospital from January 6,2020 to January 31,2022 were selected as as the study subjects,which were matched with 3 pairs(6 cases)of infertile women preparing for embryo transfer based on factors such as age,body mass index,and length of infertility.Endometrial tissue samples were collected for gene transcription and expression analysis.Twenty five women who had become pregnant through assisted reproductive technology were selected as the control group,and 28 non-pregnant women receiving ovulation monitoring at the Outpatient Department were enrolled as the case group.Status of endometrial receptivity was determined by ultrasonography.In the former group,endometrial tissues were sampled for sequencing,and GO and KEGG database enrichment analysis of differentially expressed genes was carried out.In the latter group,cervical secretions were collected,and amino acid and carnitine levels were measured by mass spectrometry.Statistical analysis was carried out using rank sum test,t test and chi-square test with SPSS v25.0 software.Results No difference was found in the clinical data of the patients with regard to age,body mass index,infertility years,AMH,FSH,LH,E2,and type of infertility.Compared with the receptive endometrial tissues,there were 100 significantly up-regulated genes and 191 significantly down-regulated genes in the pre-receptive endometrial tissue,with the most significantly altered ones being HLA-DRB5 and MMP10.The biological processes,molecular functions and pathways enriched by more differentially expressed genes in GO and KEGG were mainly immune regulation,cell adhesion and tryptophan metabolism.Analysis of secretion metabolism also revealed a significant difference in the levels of amino acids and carnitine metabolites between the two groups(P<0.05),in particular those of Alanine,Valine,3-hydroxybutyrylcarnitine(C4OH)+malonylcarnitine(C3DC)/captoylcarnitine(C10).Conclusion A significant difference has been discovered in the levels of gene transcription and protein expression in the endometrial tissues from the pre-receptive and receptive stages.The levels of amino acids and carnitine,such as Alanine,Valine,3-hydroxybutyryl carnitine(C4OH)+malonyl carnitine(C3DC)/caproyl carnitine(C10),may be associated with the receptive status of the endometrium,though this need to be verified with larger samples.