早期妊娠丢失蜕膜组织中与血管重塑相关的miRNAs表达谱分析
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摘要
目的:通过对子宫内膜蜕膜化中与血管重塑相关的miRNAs表达谱的芯片筛查及相关靶基因预测,为早期妊娠丢失的发病机制研究奠定基础。方法:利用miRNA array芯片检测早期妊娠丢失患者蜕膜组织中的差异表达miRNAs,确定与血管重塑相关的miRNAs表达谱,进而应用miRWalk2.0数据库通过生物信息学方法预测差异表达miRNAs的相关靶基因,最后应用DAVID数据库对相关靶基因进行功能富集分析(GOanalysis)及靶基因信号转导通路富集分析(KEGG Pathway analysis)。结果:早期妊娠丢失患者蜕膜组织中差异表达的miRNAs共70条,上调表达miRNAs 32条,下调表达miRNAs 38条。其中差异倍数最高(≥2.0)的2个上调miRNA为miR-125a-5p和miR-29c-3p,预测相关靶基因为血管内皮生长因子A(VEGFA)。靶基因集合功能富集于正调控RNA代谢过程、血管发育、脉管系统发育、正调控内皮细胞增殖、缺氧反应、血管生成等分子过程;KEGG通路分析涉及哺乳动物雷帕霉素靶蛋白(mTOR)信号通路、丝裂原活化蛋白激酶(MAPK)等信号通路。结论:miR-125a-5p和miR-29c-3p在早期妊娠丢失患者蜕膜组织中上调表达,可能通过对靶基因VEGFA的调控,影响子宫内膜蜕膜化血管重塑的过程,参与流产发生。
Objective:To explore the expression profile of miRNAs related to vascular remodeling of endometrium, and to predict the target genes, as a pioneer research of the pathogenesis of early pregnancy loss(EPL). Methods: The differentially expressed miRNAs in the decidual tissues of EPL were tested using the miRNA microarray, and the expression profile of miRNAs related to vascular remodeling was determined. The target genes of those differentially expressed miRNAs were screened using miRWalk2.0 database. The bioinformatics analysis of these target genes was performed by Gene Ontology analysis(GO-analysis) and Kyoto Encyclopedia of Genes and Genomes Pathway analysis(KEGG Pathway analysis). Results: There were 70 differentially expressed miRNAs in the deciduas tissues of EPL, including 32 up-regulated and 38 down-regulated miRNAs. MiR-125 a-5 p and miR-29 c-3 p were two miRNAs with the highest difference times(Ratio>2.0) in those up-regulated miRNAs, and the predicted target gene was VEGFA. The function of target gene set was mainly located in the positive regulation of RNA metabolism, vascular development, endothelial cell proliferation and hypoxia and angiogenesis. KEGG pathway analysis demonstrated that the gene set was mostly located in mTOR signaling pathway and MAPK signaling pathway. Conclusions: The up-regulated expressions of miR-125 a-5 p and miR-29 c-3 p may affect the vascular remodeling in endometrial decidualization by regulating the VEGFA expression, suggesting that miR-125 a-5 p and miR-29 c-3 p participate in the pathogenesis of EPL.
Objective:To explore the expression profile of miRNAs related to vascular remodeling of endometrium, and to predict the target genes, as a pioneer research of the pathogenesis of early pregnancy loss(EPL). Methods: The differentially expressed miRNAs in the decidual tissues of EPL were tested using the miRNA microarray, and the expression profile of miRNAs related to vascular remodeling was determined. The target genes of those differentially expressed miRNAs were screened using miRWalk2.0 database. The bioinformatics analysis of these target genes was performed by Gene Ontology analysis(GO-analysis) and Kyoto Encyclopedia of Genes and Genomes Pathway analysis(KEGG Pathway analysis). Results: There were 70 differentially expressed miRNAs in the deciduas tissues of EPL, including 32 up-regulated and 38 down-regulated miRNAs. MiR-125 a-5 p and miR-29 c-3 p were two miRNAs with the highest difference times(Ratio>2.0) in those up-regulated miRNAs, and the predicted target gene was VEGFA. The function of target gene set was mainly located in the positive regulation of RNA metabolism, vascular development, endothelial cell proliferation and hypoxia and angiogenesis. KEGG pathway analysis demonstrated that the gene set was mostly located in mTOR signaling pathway and MAPK signaling pathway. Conclusions: The up-regulated expressions of miR-125 a-5 p and miR-29 c-3 p may affect the vascular remodeling in endometrial decidualization by regulating the VEGFA expression, suggesting that miR-125 a-5 p and miR-29 c-3 p participate in the pathogenesis of EPL.
引文
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[22]Rabbani ML,Rogers PA.Role of vascular endothelial growth factor in endometrial vascular events before implantation in rats[J].Reproduction,2001,122(1):85-90.
[23]Binder NK,Evans J,Gardner DK,et al.Endometrial signals improve embryo outcome:functional role of vascular endothelial growth factor isoforms on embryo development and implantation in mice[J].Hum Reprod,2014,29(10):2278-2286.
[2]Schwenke M,Knfler M,Velicky P,et al.Control of human endometrial stromal cell motility by PDGF-BB,HB-EGF and trophoblast-secreted factors[J].PLoS One,2013,8(1):e54336.
[3]Cl-Madendag I,Madendag Y,Altinkaya S,et al.The role of VEGF and its receptors in the etiology of early pregnancy loss[J].Gynecol Endocrinol,2014,30(2):153-156.
[4]Liao XG,Li YL,Gao RF,et al.Folate deficiency decreases apoptosis of endometrium decidual cells in pregnant mice via the mitochondrial pathway[J].Nutrients,2015,7(3):1916-1932.
[5]Ventura W,Koide K,Hori K,et al.Placental expression of microRNA-17 and-19b is down-regulated in early pregnancy loss[J].Eur J Obstet Gynecol Reprod Biol,2013,169(1):28-32.
[6]Bonnal RJ,Rossi RL,Carpi D,et al.miRiadne:a web tool for consistent integration of miRNA nomenclature[J].Nucleic Acids Res,2015,43(W1):W487-W492.
[7]Friedman RC,Farh KK,Burge CB,et al.Most mammalian m RNAs are conserved targets of microRNAs[J].Genome Res,2009,19(1):92-105.
[8]Morales Prieto DM,Markert UR.MicroRNAs in pregnancy[J].JReprod Immunol,2011,88(2):106-111.
[9]Suh N,Baehner L,Moltzahn F,et al.MicroRNA function is globally suppressed in mouse oocytes and early embryos[J].Curr Biol,2010,20(3):271-277.
[10]Zhu Y,Lu H,Huo Z,et al.MicroRNA-16 inhibits feto-maternal angiogenesis and causes recurrent spontaneous abortion by targeting vascular endothelial growth factor[J].Sci Rep,2016,6:35536.
[11]Hosseini MK,Gunel T,Gumusoglu E,et al.MicroRNA expression profiling in placenta and maternal plasma in early pregnancy loss[J].Mol Med Rep,2018,17(4):4941-4952.
[12]Yang L,Engeland CG,Cheng B.Social isolation impairs oral palatal wound healing in sprague-dawley rats:a role for miR-29and miR-203 via VEGF suppression[J].PLoS One,2013,8(8):e72359.
[13]Brkic′J,Dunk C,O′Brien J,et al.MicroRNA-218-5p Promotes Endovascular Trophoblast Differentiation and Spiral Artery Remodeling[J].Mol Ther,2018,26(9):2189-2205.
[14]Braza-Bols A,Salloum-Asfar S,Marí-Alexandre J,et al.Peritoneal fluid modifies the microRNA expression profile in endometrial and endometriotic cells from women with endometriosis[J].Hum Reprod,2015,30(10):2292-2302.
[15]Che P,Liu J,Shan Z,et al.miR-125a-5p impairs endothelial cell angiogenesis in aging mice via RTEF-1 downregulation[J].Aging Cell,2014,13(5):926-934.
[16]Gong X,Tong Q,Chen Z,et al.Microvascular density and vascular endothelial growth factor and osteopontin expression during the implantation window in a controlled ovarian hyperstimulation rat model[J].Exp Ther Med,2015,9(3):773-779.
[17]Zhi Z,Yang W,Liu L.Early missed abortion is associated with villous angiogenesis via the HIF-1α/VEGF signaling pathway[J].Arch Gynecol Obstet,2018,298(3):537-543.
[18]Carmeliet P,Ferreira V,Breier G,et al.Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele[J].Nature,1996,380(6573):435-439.
[19]Ferrara N,Carver-Moore K,Chen H,et al.Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene[J].Nature,1996,380(6573):439-442.
[20]Su MT,Lin SH,Chen YC.Genetic association studies of angiogenesis-and vasoconstriction-related genes in women with recurrent pregnancy loss:a systematic review and meta-analysis[J].Hum Reprod Update,2011,17(6):803-812.
[21]Danastas K,Whittington CM,Dowland SN,et al.Ovarian Hyperstimulation Reduces Vascular Endothelial Growth Factor-ADuring Uterine Receptivity[J].Reprod Sci,2018,Jan 1,[Epub ahead of print].
[22]Rabbani ML,Rogers PA.Role of vascular endothelial growth factor in endometrial vascular events before implantation in rats[J].Reproduction,2001,122(1):85-90.
[23]Binder NK,Evans J,Gardner DK,et al.Endometrial signals improve embryo outcome:functional role of vascular endothelial growth factor isoforms on embryo development and implantation in mice[J].Hum Reprod,2014,29(10):2278-2286.