{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T15:13:51Z","timestamp":1791386031570,"version":"4.3.3"},"reference-count":46,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2023,3,21]],"date-time":"2023-03-21T00:00:00Z","timestamp":1679356800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82271734"],"award-info":[{"award-number":["82271734"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31801250"],"award-info":[{"award-number":["31801250"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2021SHRC005"],"award-info":[{"award-number":["2021SHRC005"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Social Talent Foundation of Tangdu Hospital","award":["82271734"],"award-info":[{"award-number":["82271734"]}]},{"name":"Social Talent Foundation of Tangdu Hospital","award":["31801250"],"award-info":[{"award-number":["31801250"]}]},{"name":"Social Talent Foundation of Tangdu Hospital","award":["2021SHRC005"],"award-info":[{"award-number":["2021SHRC005"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Ovarian stimulation is associated with an increased incidence of abnormal placentation. Uterine natural killer (uNK) cells are the major subpopulation of decidual immune cells, which are crucial for placentation. In a previous study, we found that ovarian stimulation impairs uNK cell density on gestation day (GD) 8.5 in mice. However, it was not clear how ovarian stimulation led to a reduction in the density of uNK cells. In this study, we constructed two mouse models, an in vitro mouse embryo transfer model and an estrogen-stimulated mouse model. We used HE and PAS glycogen staining, immunohistochemical techniques, q-PCR, Western blot, and flow cytometry to analyze the mouse decidua and placenta, and the results showed that SO resulted in a fetal weight reduction, abnormal placental morphology, decreased placental vascular density, and abnormal density and function of uNK cells. Our results suggest that ovarian stimulation resulted in aberrant estrogen signaling and may contribute to the disorder of uNK cells caused by ovarian stimulation. Together, these results provide new insights into the mechanisms of aberrant maternal endocrine environments and abnormal placentation.<\/jats:p>","DOI":"10.3390\/ijms24065907","type":"journal-article","created":{"date-parts":[[2023,3,21]],"date-time":"2023-03-21T03:15:03Z","timestamp":1679368503000},"page":"5907","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Ovarian Stimulation in Mice Resulted in Abnormal Placentation through Its Effects on Proliferation and Cytokine Production of Uterine NK Cells"],"prefix":"10.3390","volume":"24","author":[{"given":"Rong","family":"Ma","sequence":"first","affiliation":[{"name":"Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, Xi\u2019an 710038, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ni","family":"Jin","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, Xi\u2019an 710038, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Lei","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, Xi\u2019an 710038, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6786-1057","authenticated-orcid":false,"given":"Jie","family":"Dong","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, Xi\u2019an 710038, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujing","family":"Xiong","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, Xi\u2019an 710038, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenxi","family":"Qian","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, Xi\u2019an 710038, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2505-4174","authenticated-orcid":false,"given":"Shuqiang","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, Xi\u2019an 710038, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohong","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, Xi\u2019an 710038, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"103076","DOI":"10.1016\/j.ebiom.2020.103076","article-title":"Integrated multi-omics reveal epigenomic disturbance of assisted reproductive technologies in human offspring","volume":"61","author":"Chen","year":"2020","journal-title":"EBioMedicine"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1080\/03009734.2020.1726534","article-title":"Perinatal outcome in children born after assisted reproductive technologies","volume":"125","author":"Wennerholm","year":"2020","journal-title":"Ups. J. Med. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1093\/humupd\/dmz001","article-title":"The health of children conceived by ART: \u2018the chicken or the egg?\u2019","volume":"25","author":"Berntsen","year":"2019","journal-title":"Hum. Reprod. Update"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.rbmo.2017.09.009","article-title":"Perinatal outcomes after stimulated versus natural cycle IVF: A systematic review and meta-analysis","volume":"36","author":"Kamath","year":"2018","journal-title":"Reprod. Biomed. Online"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1093\/humrep\/deaa298","article-title":"Birthweight and other perinatal outcomes of singletons conceived after assisted reproduction compared to natural conceived singletons in couples with unexplained subfertility: Follow-up of two randomized clinical trials","volume":"36","author":"Wessel","year":"2021","journal-title":"Hum. Reprod."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1146\/annurev-cellbio-100814-125620","article-title":"Placenta: The forgotten organ","volume":"31","author":"Maltepe","year":"2015","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.placenta.2017.12.016","article-title":"Placental pathologic changes and perinatal outcomes in placenta previa","volume":"63","author":"Jung","year":"2018","journal-title":"Placenta"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.placenta.2020.05.003","article-title":"The placenta in fetal growth restriction: What is going wrong?","volume":"96","author":"Sun","year":"2020","journal-title":"Placenta"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1038\/s41574-020-0372-6","article-title":"Tracking placental development in health and disease","volume":"16","author":"Aplin","year":"2020","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1799","DOI":"10.1007\/s10815-018-1244-z","article-title":"Superovulation alters the expression of endometrial genes critical to tissue remodeling and placentation","volume":"35","author":"Senapati","year":"2018","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/j.rbmo.2010.04.022","article-title":"High serum oestradiol concentrations in IVF cycles increase the risk of pregnancy complications related to abnormal placentation","volume":"21","author":"Farhi","year":"2010","journal-title":"Reprod. Biomed. Online"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1093\/biolre\/iox067","article-title":"The superovulated environment, independent of embryo vitrification, results in low birthweight in a mouse model","volume":"97","author":"Weinerman","year":"2017","journal-title":"Biol. Reprod."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1095\/biolreprod.113.110411","article-title":"Peri-implantation hormonal milieu: Elucidating mechanisms of abnormal placentation and fetal growth","volume":"90","author":"Mainigi","year":"2014","journal-title":"Biol. Reprod."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2597","DOI":"10.3389\/fimmu.2018.02597","article-title":"Regulation of Placental Extravillous Trophoblasts by the Maternal Uterine Environment","volume":"9","author":"Pollheimer","year":"2018","journal-title":"Front. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1111\/imm.12218","article-title":"The up side of decidual natural killer cells: New developments in immunology of pregnancy","volume":"141","author":"Siewiera","year":"2014","journal-title":"Immunology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1016\/j.ebiom.2018.12.015","article-title":"TGFb1 suppresses the activation of distinct dNK subpopulations in preeclampsia","volume":"39","author":"Zhang","year":"2019","journal-title":"EBioMedicine"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"524","DOI":"10.1093\/biolre\/ioz203","article-title":"Natural killer cells: Functional differences in recurrent spontaneous abortiondagger","volume":"102","author":"Hao","year":"2020","journal-title":"Biol. Reprod."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"618","DOI":"10.1002\/mrd.23528","article-title":"Ovarian stimulation reduces fetal growth by dysregulating uterine natural killer cells in mice","volume":"88","author":"Zhang","year":"2021","journal-title":"Mol. Reprod. Dev."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"851","DOI":"10.1210\/er.2014-1045","article-title":"Cyclic decidualization of the human endometrium in reproductive health and failure","volume":"35","author":"Gellersen","year":"2014","journal-title":"Endocr. Rev."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1038\/nri.2017.64","article-title":"The unique immunological and microbial aspects of pregnancy","volume":"17","author":"Mor","year":"2017","journal-title":"Nat. Rev. Immunol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"R277","DOI":"10.1530\/REP-16-0453","article-title":"Sterile inflammation and pregnancy complications: A review","volume":"152","author":"Obari","year":"2016","journal-title":"Reproduction"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"8377","DOI":"10.1038\/s41598-017-07728-1","article-title":"Juxtacrine Activity of Estrogen Receptor alpha in Uterine Stromal Cells is Necessary for Estrogen-Induced Epithelial Cell Proliferation","volume":"7","author":"Winuthayanon","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.xfss.2020.06.002","article-title":"Genome-wide estrogen receptor-alpha binding and action in human endometrial stromal cells","volume":"1","author":"Yilmaz","year":"2020","journal-title":"F&S Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"764","DOI":"10.1016\/j.fertnstert.2011.06.015","article-title":"Ovarian stimulation modulates steroid receptor expression and spheroid attachment in peri-implantation endometria: Studies on natural and stimulated cycles","volume":"96","author":"Chai","year":"2011","journal-title":"Fertil. Steril."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1362","DOI":"10.1210\/me.2015-1142","article-title":"Uterine Epithelial Estrogen Receptor-alpha Controls Decidualization via a Paracrine Mechanism","volume":"29","author":"Pawar","year":"2015","journal-title":"Mol. Endocrinol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1093\/molehr\/gau065","article-title":"Ovarian stimulation using human chorionic gonadotrophin impairs blastocyst implantation and decidualization by altering ovarian hormone levels and downstream signaling in mice","volume":"20","author":"Ezoe","year":"2014","journal-title":"Mol. Hum. Reprod."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"968","DOI":"10.1016\/j.placenta.2009.08.011","article-title":"DBA-lectin reactivity defines natural killer cells that have homed to mouse decidua","volume":"30","author":"Zhang","year":"2009","journal-title":"Placenta"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1095\/biolreprod.112.102293","article-title":"DBA-lectin reactivity defines mouse uterine natural killer cell subsets with biased gene expression","volume":"87","author":"Chen","year":"2012","journal-title":"Biol. Reprod."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"960","DOI":"10.3389\/fimmu.2019.00960","article-title":"Uterine Natural Killer Cells","volume":"10","author":"Sojka","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1053\/plac.2002.0919","article-title":"Subset classification of mouse uterine natural killer cells by DBA lectin reactivity","volume":"24","author":"Paffaro","year":"2003","journal-title":"Placenta"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"6140","DOI":"10.4049\/jimmunol.181.9.6140","article-title":"Unique receptor repertoire in mouse uterine NK cells","volume":"181","author":"Yadi","year":"2008","journal-title":"J. Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1007\/978-1-60761-362-6_31","article-title":"Analysis of Uterine Natural Killer Cells in Mice","volume":"Volume 612","author":"Croy","year":"2010","journal-title":"Natural Killer Cell Protocols: Cellular and Molecular Methods"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.1016\/j.fertnstert.2012.03.028","article-title":"Peak serum estradiol level during controlled ovarian hyperstimulation is associated with increased risk of small for gestational age and preeclampsia in singleton pregnancies after in vitro fertilization","volume":"97","author":"Imudia","year":"2012","journal-title":"Fertil. Steril."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1702","DOI":"10.1016\/j.fertnstert.2007.05.036","article-title":"Single-blastocyst transfer decreases twin gestation without affecting pregnancy outcome","volume":"89","author":"Styer","year":"2008","journal-title":"Fertil. Steril."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1653","DOI":"10.1093\/hmg\/ddn055","article-title":"Superovulation alters the expression of imprinted genes in the midgestation mouse placenta","volume":"17","author":"Fortier","year":"2008","journal-title":"Hum. Mol. Genet."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1093\/humrep\/16.2.221","article-title":"The impact of ovarian stimulation on implantation and fetal development in mice","volume":"16","author":"Ertzeid","year":"2001","journal-title":"Hum. Reprod."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e31274","DOI":"10.7554\/eLife.31274","article-title":"Clearance of senescent decidual cells by uterine natural killer cells in cycling human endometrium","volume":"6","author":"Brighton","year":"2017","journal-title":"eLife"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2091","DOI":"10.1007\/s00018-019-03395-9","article-title":"Epigenetic modifications working in the decidualization and endometrial receptivity","volume":"77","author":"Liu","year":"2020","journal-title":"Cell Mol. Life Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"6760","DOI":"10.4049\/jimmunol.173.11.6760","article-title":"Recruitment of uterine NK cells: Induction of CXC chemokine ligands 10 and 11 in human endometrium by estradiol and progesterone","volume":"173","author":"Sentman","year":"2004","journal-title":"J. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"664","DOI":"10.1016\/j.bbrc.2013.04.106","article-title":"Genetic deletion of Cxcl14 in mice alters uterine NK cells","volume":"435","author":"Cao","year":"2013","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1186\/s12958-017-0290-1","article-title":"The regulation of ovary and conceptus on the uterine natural killer cells during early pregnancy","volume":"15","author":"Gong","year":"2017","journal-title":"Reprod. Biol. Endocrinol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"eabb7800","DOI":"10.1126\/sciimmunol.abb7800","article-title":"Continuous human uterine NK cell differentiation in response to endometrial regeneration and pregnancy","volume":"6","author":"Strunz","year":"2021","journal-title":"Sci. Immunol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"4267","DOI":"10.4049\/jimmunol.178.7.4267","article-title":"Genetic deletion of placenta growth factor in mice alters uterine NK cells","volume":"178","author":"Tayade","year":"2007","journal-title":"J. Immunol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"12542","DOI":"10.1073\/pnas.0901647106","article-title":"De novo synthesis of estrogen in pregnant uterus is critical for stromal decidualization and angiogenesis","volume":"106","author":"Das","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"9596","DOI":"10.1074\/jbc.RA120.012753","article-title":"The transcription factor HAND2 up-regulates transcription of the IL15 gene in human endometrial stromal cells","volume":"295","author":"Murata","year":"2020","journal-title":"J. Biol. Chem."},{"key":"ref_46","first-page":"e55168","article-title":"Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization","volume":"121","author":"Hennes","year":"2017","journal-title":"J. Vis. Exp."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/6\/5907\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:59:26Z","timestamp":1760122766000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/6\/5907"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,21]]},"references-count":46,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2023,3]]}},"alternative-id":["ijms24065907"],"URL":"https:\/\/doi.org\/10.3390\/ijms24065907","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,21]]}}}