PMID- 28122716 OWN - NLM STAT- MEDLINE DCOM- 20170627 LR - 20230802 IS - 1522-1490 (Electronic) IS - 0363-6119 (Print) IS - 0363-6119 (Linking) VI - 312 IP - 4 DP - 2017 Apr 1 TI - Relaxin confers cytotrophoblast protection from hypoxia-reoxygenation injury through the phosphatidylinositol 3-kinase-Akt/protein kinase B cell survival pathway. PG - R559-R568 LID - 10.1152/ajpregu.00306.2016 [doi] AB - Preeclampsia is a hypertensive syndrome that manifests after 20 wk of gestation. Contemporary understanding of the maternal-fetal interface in preeclampsia suggests a major role for placental oxidative stress resulting from ischemia-reperfusion injury. We hypothesized that the pregnancy hormone relaxin would reduce cytotrophoblast apoptosis and necrosis (aponecrosis) and, hence, the export of placental debris into the maternal circulation. If so, then relaxin might be employed as a therapeutic intervention to diminish the activation of the maternal systemic inflammatory response central to the development of clinical disease. HTR-8/SVneo cells, a model for first trimester extravillous trophoblast, were subjected to serum deprivation and hypoxia or hypoxia-reoxygenation. The cells were treated with recombinant human relaxin or vehicle and apoptosis and/or necrosis evaluated by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), CellEvent Caspase-3/7 and SYTOX AADvanced kit, and propidium iodide staining as determined by fluorescence microscopy or flow cytometry. To interrogate mechanisms of relaxin cytoprotection, HTR-8/SVneo cells were pretreated with pharmacological inhibitors of PI3-kinase LY294004, Akt/PKB MK-2206, or DMSO vehicle. HTR-8/SVneo cell identity was first confirmed by RT-PCR. The cells expressed placental alkaline phosphatase, aromatase, and human leukocyte antigen G. In addition, the cells expressed the relaxin receptor RXFP1 as well as H1 and H2 relaxins. Serum deprivation and hypoxia increased apoptotic cell death in HTR-8/SVneo cells, which was significantly ameliorated by concurrent treatment with relaxin. Serum deprivation and hypoxia-reoxygenation increased necrotic cell death in HTR-8/SVneo cells, which was also significantly rescued by concurrent treatment with relaxin. Pretreatment with LY294002 or MK-2206, to inhibit the phosphatidylinositol 3-kinase-Akt/protein kinase B cell survival pathway, significantly blunted the cytoprotective effect of relaxin. We demonstrated trophoblast cytoprotection by intervention with supraphysiological concentrations of relaxin, a process in part mediated through the PI3-kinase-Akt/PKB cell survival pathway. These results provide further rationale for clinical investigation of relaxin as a potential therapeutic in preeclampsia. CI - Copyright (c) 2017 the American Physiological Society. FAU - Ogunleye, Oluseyi AU - Ogunleye O AD - Department of Obstetrics and Gynecology, D. H. Barron Reproductive and Perinatal Biology Research Program, University of Florida College of Medicine, Gainesville, Florida; and. FAU - Campo, Bertha AU - Campo B AD - Department of Physiology and Functional Genomics, D. H. Barron Reproductive and Perinatal Biology Research Program, University of Florida College of Medicine, Gainesville, Florida. FAU - Herrera, Diana AU - Herrera D AD - Department of Physiology and Functional Genomics, D. H. Barron Reproductive and Perinatal Biology Research Program, University of Florida College of Medicine, Gainesville, Florida. FAU - Post Uiterweer, Emiel D AU - Post Uiterweer ED AD - Department of Physiology and Functional Genomics, D. H. Barron Reproductive and Perinatal Biology Research Program, University of Florida College of Medicine, Gainesville, Florida. FAU - Conrad, Kirk P AU - Conrad KP AD - Department of Obstetrics and Gynecology, D. H. Barron Reproductive and Perinatal Biology Research Program, University of Florida College of Medicine, Gainesville, Florida; and kpconrad@ufl.edu. AD - Department of Physiology and Functional Genomics, D. H. Barron Reproductive and Perinatal Biology Research Program, University of Florida College of Medicine, Gainesville, Florida. LA - eng GR - R25 HL103181/HL/NHLBI NIH HHS/United States GR - P01 HD065647/HD/NICHD NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20170125 PL - United States TA - Am J Physiol Regul Integr Comp Physiol JT - American journal of physiology. Regulatory, integrative and comparative physiology JID - 100901230 RN - 9002-69-1 (Relaxin) RN - EC 2.7.1.137 (Phosphatidylinositol 3-Kinase) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) SB - IM MH - B-Lymphocytes/*drug effects/pathology MH - Cell Line MH - Cell Survival/*drug effects MH - Cytoprotection/drug effects MH - Humans MH - Phosphatidylinositol 3-Kinase/*metabolism MH - Proto-Oncogene Proteins c-akt/metabolism MH - Relaxin/*administration & dosage/metabolism MH - Reperfusion Injury/*metabolism/prevention & control MH - Signal Transduction/drug effects MH - Trophoblasts/drug effects/*metabolism/pathology PMC - PMC5407077 OTO - NOTNLM OT - H2 relaxin OT - HTR-8/SVneo cell OT - apoptosis OT - hypoxia OT - hypoxia-reoxygenation OT - necrosis OT - preeclampsia EDAT- 2017/01/27 06:00 MHDA- 2017/06/28 06:00 PMCR- 2018/04/01 CRDT- 2017/01/27 06:00 PHST- 2016/07/07 00:00 [received] PHST- 2017/01/04 00:00 [revised] PHST- 2017/01/20 00:00 [accepted] PHST- 2017/01/27 06:00 [pubmed] PHST- 2017/06/28 06:00 [medline] PHST- 2017/01/27 06:00 [entrez] PHST- 2018/04/01 00:00 [pmc-release] AID - ajpregu.00306.2016 [pii] AID - R-00306-2016 [pii] AID - 10.1152/ajpregu.00306.2016 [doi] PST - ppublish SO - Am J Physiol Regul Integr Comp Physiol. 2017 Apr 1;312(4):R559-R568. doi: 10.1152/ajpregu.00306.2016. Epub 2017 Jan 25.