title: GPR30 decreases cardiac chymase/angiotensin II by inhibiting local mast cell number.
abstract: Chronic activation of the novel estrogen receptor GPR30 by its agonist G1 mitigates the adverse effects of estrogen (E2) loss on cardiac structure and function. Using the ovariectomized (OVX) mRen2.Lewis rat, an E2-sensitive model of diastolic dysfunction, we found that E2 status is inversely correlated with local cardiac angiotensin II (Ang II) levels, likely via Ang I/chymase-mediated production. Since chymase is released from cardiac mast cells during stress (e.g., volume/pressure overload, inflammation), we hypothesized that GPR30-related cardioprotection after E2 loss might occur through its opposing actions on cardiac mast cell proliferation and chymase production. Using real-time quantitative PCR, immunohistochemistry, and immunoblot analysis, we found mast cell number, chymase expression, and cardiac Ang II levels were significantly increased in the hearts of OVX-compared to ovary-intact mRen2.Lewis rats and the GPR30 agonist G1 (50 mg/kg/day, s.c.) administered for 2 weeks limited the adverse effects of estrogen loss. In vitro studies revealed that GPR30 receptors are expressed in the RBL-2H3 mast cell line and G1 inhibits serum-induced cell proliferation in a dose-dependent manner, as determined by cell counting, BrdU incorporation assay, and Ki-67 staining. Using specific antagonists to estrogen receptors, blockage of GPR30, but not ERα or ERβ, attenuated the inhibitory effects of estrogen on BrdU incorporation in RBL-2H3 cells. Further study of the mechanism underlying the effect on cell proliferation showed that G1 inhibits cyclin-dependent kinase 1 (CDK1) mRNA and protein expression in RBL-2H3 cells in a dose-dependent manner.
authors.affiliation: Department of Anesthesiology, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27159-1009, USA; Department of Cardiology, Jinan Central Hospital, Affiliated with Shandong University, 105 Jiefang Road, Jinan, 250013, China.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: Department of Anesthesiology, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27159-1009, USA.
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Anesthesiology, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27159-1009, USA.
authors.forename: Marina
authors.initials: M
authors.lastname: Lin
authors.affiliation: Department of Anesthesiology, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27159-1009, USA; Hypertension and Vascular Disease Center, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA; Office of Women in Medicine and Science, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA. Electronic address: lgroban@wakehealth.edu.
authors.forename: Leanne
authors.initials: L
authors.lastname: Groban
mesh_terms: D000804:Angiotensin II / Q000378:metabolism*; D000818:Animals; D016203:CDC2 Protein Kinase; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D053818:Chymases / Q000378:metabolism*; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D003517:Cyclopentanes / Q000494:pharmacology; D004958:Estradiol / Q000378:metabolism / Q000494:pharmacology; D065171:Estrogen Receptor Antagonists / Q000494:pharmacology; D004967:Estrogens / Q000378:metabolism; D005260:Female; D008407:Mast Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D009206:Myocardium / Q000166:cytology / Q000378:metabolism; D010052:Ovariectomy; D011804:Quinolines / Q000494:pharmacology; D011917:Rats, Inbred Lew; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000378:metabolism*
chemical_list: C517943:1-(4-(6-bromobenzo(1,3)dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta(c)quinolin-8-yl)ethanone; D003517:Cyclopentanes; D065171:Estrogen Receptor Antagonists; D004967:Estrogens; C520305:Gper1 protein, rat; D011804:Quinolines; D043562:Receptors, G-Protein-Coupled; D000804:Angiotensin II; D004958:Estradiol; D016203:CDC2 Protein Kinase; C548796:Cdk1 protein, rat; D018844:Cyclin-Dependent Kinases; D053818:Chymases
keywords: Chymase; Estrogen; GPR30; Heart; Mast cell