miR-146a-5p在不明原因复发性流产蜕膜组织中的表达及其意义

doi:10.12280/gjszjk.20200669

摘要/Abstract

摘要：

目的: 探讨miR-146a-5p在不明原因复发性流产（URSA）蜕膜组织中的表达变化并分析其可能作用机制。方法: 选取2019年1—12月在成都西囡妇科医院因URSA行清宫术患者（URSA组）及因无生育计划行人工流产术的正常妊娠者（正常妊娠组）各10例,收集2组所有对象的蜕膜组织。比较2组蜕膜组织中miR-146a-5p水平,利用生物信息学方法预测miR-146a-5p潜在靶基因和相关分子机制,并检测蜕膜组织中关键靶分子水平并分析其与miR-146a-5p水平之间的相关性。结果: URSA组miR-146a-5p水平低于正常妊娠组（P<0.001）。在至少3个数据库均预测到的miR-146a-5p靶基因共35个,其中IRAK1、CD80和TRAF6富集于Toll样受体（TLR）信号通路;IRAK1、SORT1和TRAF6富集于神经营养素（NT）信号通路,miR-146a-5p预测靶基因编码的功能蛋白之间存在3对相互作用关系。URSA组IRAK1、CD80、TRAF6和SORT1基因和蛋白水平均高于正常妊娠组（P<0.05）,miR-146a-5p水平与这4个分子基因和蛋白水平均呈负相关（P<0.05）。结论: 蜕膜组织中miR-146a-5p可能通过调控TLR和NT信号通路参与URSA发病机制,为后续URSA防治策略研究提供了新的思路和靶点。

关键词: 流产, 习惯性, 蜕膜, MicroRNA-146a-5p, Toll样受体, 神经生长因子类, 信号传导

Abstract:

Objective: To study the expression of miR-146a-5p in the decidual tissue of unexplained recurrent miscarriage (URSA), and analyze the possible mechanism of action. Methods: From January to December 2019, ten patients who underwent uterine evacuation due to URSA (URSA group) and ten normal pregnant patients who underwent induced abortion due to no childbirth planning (control group) in Chengdu Xi′nan Gynaecology Hospital were recruited. The decidual tissues were collected. The levels of miR-146a-5p in the decidual tissues were tested. Bioinformatics methods were used to predict the potential target genes of miR-146a-5p and the molecular mechanisms involved. The levels of key target genes of miR-146a-5p in each group were detected. The correlation between miR-146a-5p and the target genes was analyzed.Results: The level of miR-146a-5p in decidual tissue was lower in the URSA group than in the control group (P<0.001). Thirty-five target genes of miR-146a-5p were predicted in at least three databases, among whichIRAK1, CD80, and TRAF6 in the Toll-like receptor (TLR) signaling pathway; and IRAK1, SORT1, and TRAF6 in the neurotrophin (NT) signaling pathway. The levels of IRAK1, CD80, TRAF6 and SORT1, both mRNAs and proteins, in decidual tissue in the URSA group were higher than those in the control group (all P<0.05), and the level of miR-146a-5p was negatively correlated with the expressions of these four molecules (P<0.05).Conclusions: miR-146a-5p in decidual tissue may participate in the pathogenesis of URSA by regulating TLR and NT signaling pathways, which provids new ideas and potential targets for further studies on URSA prevention and therapy.

Key words: Abortion, habitual, Decidua, MicroRNA-146a-5p, Toll-like receptors, Nerve growth factors, Signal transduction

冯玉婷, 罗姝红, 李兰, 苏琴, 程惊秋, 叶红霞. miR-146a-5p在不明原因复发性流产蜕膜组织中的表达及其意义[J]. 国际生殖健康/计划生育, 2021, 40(4): 286-293.

FENG Yu-ting, LUO Shu-hong, LI Lan, SU Qin, CHENG Jing-qiu, YE Hong-xia. Expression of MiR-146a-5p in Decidual Tissue of Unexplained Recurrent Spontaneous Abortion[J]. Journal of International Reproductive Health/Family Planning, 2021, 40(4): 286-293.

图/表 13

参考文献 28

[1]	Toth B, Würfel W, Bohlmann M, et al. Recurrent Miscarriage: Diagnostic and Therapeutic Procedures. Guideline of the DGGG, OEGGG and SGGG (S2k-Level, AWMF Registry Number 015/050)[J]. Geburtshilfe Frauenheilkd, 2018, 78(4):364-381. doi: 10.1055/a-0586-4568. doi: 10.1055/a-0586-4568 URL
[2]	Muyayalo KP, Li ZH, Mor G, et al. Modulatory effect of intravenous immunoglobulin on Th17/Treg cell balance in women with unexplained recurrent spontaneous abortion[J]. Am J Reprod Immunol, 2018, 80(4):e13018. doi: 10.1111/aji.13018. doi: 10.1111/aji.13018 URL
[3]	自然流产诊治中国专家共识编写组. 自然流产诊治中国专家共识(2020年版)[J]. 中国实用妇科与产科杂志, 2020, 36(11):1082-1090. doi: 10.19538/j.fk2020110113. doi: 10.19538/j.fk2020110113
[4]	Konečná B, Lauková L, Vlková B. Immune activation by nucleic acids: A role in pregnancy complications[J]. Scand J Immunol, 2018, 87(4):e12651. doi: doi: 10.1111/sji.12651. doi: doi: 10.1111/sji.12651 URL
[5]	李丽斐, 李发敏子, 吕萌, 等. 宫颈炎及其临床治疗对妊娠结局的影响[J]. 中华生殖与避孕杂志, 2020, 40(6):511-514. doi: 10.3760/cma.j.cn101441-20190525-00205. doi: 10.3760/cma.j.cn101441-20190525-00205
[6]	秦诗, 高玉平. 复发性流产的免疫学因素研究进展[J]. 中华生殖与避孕杂志, 2018, 38(2):165-169. doi: 10.3760/cma.j.issn.2096-2916.2018.02.015. doi: 10.3760/cma.j.issn.2096-2916.2018.02.015
[7]	俞田田, 洪丽华. 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[J]. 国际生殖健康/计划生育杂志, 2019, 38(1):43-47. doi: 10.3969/j.issn.1674-1889.2019.01.008. doi: 10.3969/j.issn.1674-1889.2019.01.008
[8]	Fogel O, Bugge Tinggaard A, Fagny M, et al. Deregulation of microRNA expression in monocytes and CD4(+) T lymphocytes from patients with axial spondyloarthritis[J]. Arthritis Res Ther, 2019, 21(1):51. doi: 10.1186/s13075-019-1829-7. doi: 10.1186/s13075-019-1829-7 URL
[9]	Hou T, Liao J, Zhang C, et al. Elevated expression of miR-146, miR-139 and miR-340 involved in regulating Th1/Th2 balance with acute exposure of fine particulate matter in mice[J]. Int Immunopharmacol, 2018, 54:68-77. doi: 10.1016/j.intimp.2017. 10.003. doi: 10.1016/j.intimp.2017. 10.003 URL
[10]	Zhao L, Li J, Huang S. Patients with Unexplained Recurrent Spontaneous Abortion Show Decreased Levels of Microrna-146a-5p in the Deciduae[J]. Ann Clin Lab Sci, 2018, 48(2):177-182.
[11]	马亚玲, 刘建国. 复发性流产病因及治疗进展[J]. 辽宁中医药大学学报, 2020, 22(7):117-122. doi: 10.13194/j.issn.1673-842x.2020.07.028. doi: 10.13194/j.issn.1673-842x.2020.07.028
[12]	McCulloch K, Litherland GJ, Rai TS. Cellular senescence in osteoarthritis pathology[J]. Aging Cell, 2017, 16(2):210-218. doi: 10.1111/acel.12562. doi: 10.1111/acel.12562 pmid: 28124466
[13]	孟楠, 张璇. MicroRNAs在复发性流产病理过程中作用的研究进展[J]. 中华生殖与避孕杂志, 2019, 39(11):921-925. doi: 10.3760/cma.j.issn.2096-2916.2019.11.011. doi: 10.3760/cma.j.issn.2096-2916.2019.11.011
[14]	Zhang Y, Zhou J, Li MQ, et al. MicroRNA-184 promotes apoptosis of trophoblast cells via targeting WIG1 and induces early spontaneous abortion[J]. Cell Death Dis, 2019, 10(3):223. doi: 10.1038/s41419-019-1443-2. doi: 10.1038/s41419-019-1443-2 pmid: 30833572
[15]	Zhu X, Liu H, Zhang Z, et al. MiR-103 protects from recurrent spontaneous abortion via inhibiting STAT1 mediated M1 macrophage polarization[J]. Int J Biol Sci, 2020, 16(12):2248-2264. doi: 10.7150/ijbs.46144. doi: 10.7150/ijbs.46144 URL
[16]	Huang Q, Ding J, Gong M, et al. Effect of miR-30e regulating NK cell activities on immune tolerance of maternal-fetal interface by targeting PRF1[J]. Biomed Pharmacother, 2019, 109:1478-1487. doi: 10.1016/j.biopha.2018.09.172. doi: S0753-3322(18)33170-6 pmid: 30551399
[17]	Bae SC, Lee YH. MiR-146a levels in rheumatoid arthritis and their correlation with disease activity: a meta-analysis[J]. Int J Rheum Dis, 2018, 21(7):1335-1342. doi: 10.1111/1756-185X.13338. doi: 10.1111/1756-185X.13338 URL
[18]	Shomali N, Mansoori B, Mohammadi A, et al. MiR-146a functions as a small silent player in gastric cancer[J]. Biomed Pharmacother, 2017, 96:238-245. doi: 10.1016/j.biopha.2017.09.138. doi: S0753-3322(17)33369-3 pmid: 28987948
[19]	Kim SY, Heo S, Kim SH, et al. Suppressive effects of dehydrocostus lactone on the toll-like receptor signaling pathways[J]. Int Immunopharmacol, 2020, 78:106075. doi: 10.1016/j.intimp.2019. 106075. doi: 10.1016/j.intimp.2019. 106075 URL
[20]	Noh JY, Yoon SR, Kim TD, et al. Toll-Like Receptors in Natural Killer Cells and Their Application for Immunotherapy[J]. J Immunol Res, 2020, 2020:2045860. doi: 10.1155/2020/2045860. doi: 10.1155/2020/2045860
[21]	MacLeod H, Wetzler LM. T cell activation by TLRs: a role for TLRs in the adaptive immune response[J]. Sci STKE, 2007, 2007(402):pe48. doi: 10.1126/stke.4022007pe48. doi: 10.1126/stke.4022007pe48
[22]	Hochrein H, O′Keeffe M, Wagner H. Human and mouse plasmacytoid dendritic cells[J]. Hum Immunol, 2002, 63(12):1103-1110. doi: 10.1016/s0198-8859(02)00748-6. doi: 10.1016/s0198-8859(02)00748-6 URL
[23]	Nahid MA, Satoh M, Chan EK. Mechanistic role of microRNA-146a in endotoxin-induced differential cross-regulation of TLR signaling[J]. J Immunol, 2011, 186(3):1723-1734. doi: 10.4049/jimmunol.1002311. doi: 10.4049/jimmunol.1002311 pmid: 21178010
[24]	Hung PS, Liu CJ, Chou CS, et al. miR-146a enhances the oncogenicity of oral carcinoma by concomitant targeting of the IRAK1, TRAF6 and NUMB genes[J]. PLoS One, 2013, 8(11):e79926. doi: 10.1371/journal.pone.0079926. doi: 10.1371/journal.pone.0079926 URL
[25]	Park H, Huang X, Lu C, et al. MicroRNA-146a and microRNA-146b regulate human dendritic cell apoptosis and cytokine production by targeting TRAF6 and IRAK1 proteins[J]. J Biol Chem, 2015, 290(5):2831-2841. doi: 10.1074/jbc.M114.591420. doi: 10.1074/jbc.M114.591420 URL
[26]	Tometten M, Blois S, Kuhlmei A, et al. Nerve growth factor translates stress response and subsequent murine abortion via adhesion molecule-dependent pathways[J]. Biol Reprod, 2006, 74(4):674-683. doi: 10.1095/biolreprod.105.044651. doi: 10.1095/biolreprod.105.044651 pmid: 16371592
[27]	Ramer I, Kruczek A, Doulaveris G, et al. Reduced Circulating Concentration of Brain-derived Neurotrophic Factor is Associated with Peri- and Post-implantation Failure following In Vitro Fertilization-Embryo Transfer[J]. Am J Reprod Immunol, 2016, 75(1):36-41. doi: 10.1111/aji.12430. doi: 10.1111/aji.12430 URL
[28]	Vanmierlo T, De Vry J, Nelissen E, et al. Gestational stress in mouse dams negatively affects gestation and postpartum hippocampal BDNF and P11 protein levels[J]. Mol Cell Neurosci, 2018, 88:292-299. doi: 10.1016/j.mcn.2018.02.009. doi: 10.1016/j.mcn.2018.02.009 URL

基因	引物序列	引物大小（μg/μmol）
IRAK1	上游：GAGAGTGACGAGAGCCTAGG	6 256.13
	下游：CTCGATTCTCCTGCCGTGTC	6 010.94
CD80	上游：GTGGTCACAATGTTTCTGTTGA	6 771.46
	下游：GTTCTTGTACTCGGGCCATATA	6 716.42
TRAF6	上游：GAGACAGGTTTCTTGTGACAAC	6 774.47
	下游：TGGCAACCAAAAGTACTGAATG	6 776.50
SORT1	上游：TCTCAGAGCCGAATGCCGTAGG	6 760.45
	下游：GCCACAATGATGCCTCCAGAATCC	7 266.80

基因	引物序列	引物大小（μg/μmol）
IRAK1	上游：GAGAGTGACGAGAGCCTAGG	6 256.13
	下游：CTCGATTCTCCTGCCGTGTC	6 010.94
CD80	上游：GTGGTCACAATGTTTCTGTTGA	6 771.46
	下游：GTTCTTGTACTCGGGCCATATA	6 716.42
TRAF6	上游：GAGACAGGTTTCTTGTGACAAC	6 774.47
	下游：TGGCAACCAAAAGTACTGAATG	6 776.50
SORT1	上游：TCTCAGAGCCGAATGCCGTAGG	6 760.45
	下游：GCCACAATGATGCCTCCAGAATCC	7 266.80

组别	n	年龄（岁）	胎龄（d）	流产家族史	流产次数（次）	蜕膜组织miR- 146a-5p水平
正常妊娠组	10	28.70±4.79	52.40±5.36	0（0）	0	1.01±0.16
URSA组	10	32.50±3.75	57.10±5.76	1（0.10）	2.30±0.48	0.47±0.17
t或χ²		1.977	1.889	1.053	-	7.452
P		0.064	0.075	0.305	-	<0.001

组别	n	年龄（岁）	胎龄（d）	流产家族史	流产次数（次）	蜕膜组织miR- 146a-5p水平
正常妊娠组	10	28.70±4.79	52.40±5.36	0（0）	0	1.01±0.16
URSA组	10	32.50±3.75	57.10±5.76	1（0.10）	2.30±0.48	0.47±0.17
t或χ²		1.977	1.889	1.053	-	7.452
P		0.064	0.075	0.305	-	<0.001

基因ID	名称	序列
MIMAT0000449	hsa-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUU
MIMAT0000158	mmu-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUU
MIMAT0000852	rno-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUU
MIMAT0001163	gga-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUU
MIMAT0006203	mml-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUU
MIMAT0007230	oan-miR-146a-5p	UGAGAACUGAAUUCCACGGGUU
MIMAT0012746	mdo-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUU
MIMAT0014622	tgu-miR-146a-5p	UGAGAACUGAAUUCCAUGGACU
MIMAT0021775	aca-miR-146a-5p	UGAGAACUGAAUUCCAUAGGC
MIMAT0022635	ola-miR-146a-5p	UGAGAACUGAAUUCCAUAGAUGGUA
MIMAT0022963	ssc-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUU
MIMAT0032371	ssa-miR-146a-5p	UGAGAACUGAAUUCCAUAGAUGG
MIMAT0036567	tch-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUU
MIMAT0037774	cpi-miR-146a-5p	UGAGAACUGAAUUCCAUGUGUUG
MIMAT0038171	ami-miR-146a-5p	UGAGAACUGAAUUCCAUGGGU
MIMAT0038535	cli-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUUG
MIMAT0038961	pbv-miR-146a-5p	UGAGAACUGAAUUCCAUAGGCUU
MIMAT0040181	pal-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUU
MIMAT0046462	xla-miR-146a-5p	UGAGAACUGAAUUCCAUAGGU
MIMAT0047016	cpo-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUUG
MIMAT0047663	dno-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUUU
MIMAT0048253	ocu-miR-146a-5p	UGAGAACUGAAUUCCAUGGGUUG