PMID- 28437005 OWN - NLM STAT- MEDLINE DCOM- 20180417 LR - 20200403 IS - 1469-7793 (Electronic) IS - 0022-3751 (Print) IS - 0022-3751 (Linking) VI - 595 IP - 14 DP - 2017 Jul 15 TI - Convergent ERK1/2, p38 and JNK mitogen activated protein kinases (MAPKs) signalling mediate catecholoestradiol-induced proliferation of ovine uterine artery endothelial cells. PG - 4663-4676 LID - 10.1113/JP274119 [doi] AB - KEY POINTS: The catechol metabolites of 17beta-oestradiol (E(2) beta), 2-hydroxyoestradiol (2-OHE(2) ) and 4-hydroxyoestradiol (4-OHE(2) ), stimulate proliferation of pregnancy-derived ovine uterine artery endothelial cells (P-UAECs) through beta-adrenoceptors (beta-ARs) and independently of the classic oestrogen receptors (ERs). Herein we show that activation of ERK1/2, p38 and JNK mitogen activated protein kinases (MAPKs) is necessary for 2-OHE(2) - and 4-OHE(2) -induced P-UAEC proliferation, as well as proliferation induced by the parent hormone E(2) beta and other beta-AR signalling hormones (i.e. catecholamines). Conversely, although 2-OHE(2) and 4-OHE(2) rapidly activate phosphatidylinositol 3-kinase (PI3K), its activation is not involved in catecholoestradiol-induced P-UAEC proliferation. We also show for the first time the signalling mechanisms involved in catecholoestradiol-induced P-UAEC proliferation; which converge at the level of MAPKs with the signalling mechanisms mediating E(2) beta- and catecholamine-induced proliferation. The present study advances our understanding of the complex signalling mechanisms involved in regulating uterine endothelial cell proliferation during pregnancy. ABSTRACT: Previously we demonstrated that the biologically active metabolites of 17beta-oestradiol, 2-hydroxyoestradiol (2-OHE(2) ) and 4-hydroxyoestradiol (4-OHE(2) ), stimulate pregnancy-specific proliferation of uterine artery endothelial cells derived from pregnant (P-UAECs), but not non-pregnant ewes. However, unlike 17beta-oestradiol, which induces proliferation via oestrogen receptor-beta (ER-beta), the catecholoestradiols mediate P-UAEC proliferation via beta-adrenoceptors (beta-AR) and independently of classic oestrogen receptors. Herein, we aim to further elucidate the signalling mechanisms involved in proliferation induced by catecholoestradiols in P-UAECs. P-UAECs were treated with 2-OHE(2) and 4-OHE(2) for 0, 0.25, 0.5, 1, 2, 4, 12 and 24 h, to analyse activation of mitogen activated protein kinases (MAPKs) and phosphatidylinositol 3-kinase (PI3K)-AKT. Specific inhibitors for ERK1/2 MAPK (PD98059), p38 MAPK (SB203580), JNK MAPK (SP600125), or PI3K (LY294002) were used to determine the involvement of individual kinases in agonist-induced P-UAEC proliferation. 2-OHE(2) and 4-OHE(2) stimulated biphasic phosphorylation of ERK1/2, slow p38 and JNK phosphorylation over time, and rapid monophasic AKT phosphorylation. Furthermore, ERK1/2, p38 and JNK MAPKs, but not PI3K, were individually necessary for catecholoestradiol-induced proliferation. In addition, when comparing the signalling mechanisms of the catecholoestradiols, to 17beta-oestradiol and catecholamines, we observed that convergent MAPKs signalling pathways facilitate P-UAEC proliferation induced by all of these hormones. Thus, all three members of the MAPK family mediate the mitogenic effects of catecholoestradiols in the endothelium during pregnancy. Furthermore, the convergent signalling of MAPKs involved in catecholoestradiol-, 17beta-oestradiol- and catecholamine-induced endothelial cell proliferation may be indicative of unappreciated evolutionary functional redundancy to facilitate angiogenesis and ensure maintenance of uterine blood flow during pregnancy. CI - (c) 2017 University of South Florida. The Journal of Physiology (c) 2017 The Physiological Society. FAU - Landeros, Rosalina Villalon AU - Landeros RV AD - Department of Obstetrics and Gynaecology, Perinatal Research Laboratories, University of Wisconsin-Madison, Madison, WI, USA. FAU - Jobe, Sheikh O AU - Jobe SO AD - Department of Obstetrics and Gynaecology, Perinatal Research Laboratories, University of Wisconsin-Madison, Madison, WI, USA. FAU - Aranda-Pino, Gabrielle AU - Aranda-Pino G AD - Department of Obstetrics and Gynaecology, Perinatal Research Laboratories, University of Wisconsin-Madison, Madison, WI, USA. FAU - Lopez, Gladys E AU - Lopez GE AD - Department of Obstetrics and Gynaecology, Perinatal Research Laboratories, University of Wisconsin-Madison, Madison, WI, USA. FAU - Zheng, Jing AU - Zheng J AUID- ORCID: 0000-0002-2224-2372 AD - Department of Obstetrics and Gynaecology, Perinatal Research Laboratories, University of Wisconsin-Madison, Madison, WI, USA. FAU - Magness, Ronald R AU - Magness RR AUID- ORCID: 0000-0003-4680-5069 AD - Department of Obstetrics and Gynaecology, Perinatal Research Laboratories, University of Wisconsin-Madison, Madison, WI, USA. AD - Department of Pediatrics and Animal Sciences, University of Wisconsin-Madison, Madison, WI, USA. AD - Department of Animal Sciences, University of Wisconsin-Madison, Madison, WI, USA. AD - Department of Obstetrics and Gynaecology, University of South Florida Perinatal Research Vascular Centre, Morsani College of Medicine, Tampa, FL, USA. LA - eng GR - R25 GM083252/GM/NIGMS NIH HHS/United States GR - T32 HD041921/HD/NICHD NIH HHS/United States GR - R01 HL087144/HL/NHLBI NIH HHS/United States GR - F31 HD088096/HD/NICHD NIH HHS/United States GR - P01 HD038843/HD/NICHD NIH HHS/United States GR - R01 HL049210/HL/NHLBI NIH HHS/United States GR - R01 HL117341/HL/NHLBI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural DEP - 20170605 PL - England TA - J Physiol JT - The Journal of physiology JID - 0266262 RN - 0 (Estrogens, Catechol) RN - 0 (Protein Kinase Inhibitors) RN - 4TI98Z838E (Estradiol) RN - AYU2L67YUU (2-hydroxyestradiol) RN - C3ZO03450E (4-hydroxyestradiol) RN - EC 2.7.1.- (Phosphatidylinositol 3-Kinases) RN - EC 2.7.11.24 (Mitogen-Activated Protein Kinases) SB - IM CIN - J Physiol. 2017 Jul 15;595(14):4571-4572. PMID: 28513857 EIN - J Physiol. 2017 Nov 1;595(21):6789. PMID: 29090470 MH - Animals MH - Cell Proliferation/drug effects MH - Cells, Cultured MH - Endothelial Cells/*drug effects/metabolism MH - Estradiol/*analogs & derivatives/pharmacology MH - Estrogens, Catechol/*pharmacology MH - Female MH - Mitogen-Activated Protein Kinases/*metabolism MH - Phosphatidylinositol 3-Kinases/metabolism MH - Protein Kinase Inhibitors/pharmacology MH - Sheep MH - Signal Transduction MH - Uterine Artery/*cytology PMC - PMC5509880 OTO - NOTNLM OT - MAPKs OT - endothelial cell OT - oestrogens EDAT- 2017/04/25 06:00 MHDA- 2018/04/18 06:00 PMCR- 2018/07/15 CRDT- 2017/04/25 06:00 PHST- 2017/02/01 00:00 [received] PHST- 2017/04/10 00:00 [accepted] PHST- 2017/04/25 06:00 [pubmed] PHST- 2018/04/18 06:00 [medline] PHST- 2017/04/25 06:00 [entrez] PHST- 2018/07/15 00:00 [pmc-release] AID - TJP12388 [pii] AID - 10.1113/JP274119 [doi] PST - ppublish SO - J Physiol. 2017 Jul 15;595(14):4663-4676. doi: 10.1113/JP274119. Epub 2017 Jun 5.