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Activation of GPR30 inhibits cardiac fibroblast proliferation

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Abstract

The incidence of left ventricular diastolic dysfunction significantly increases in postmenopausal women suggesting the association between estrogen loss and diastolic dysfunction. The in vivo activation of G protein-coupled estrogen receptor (GPR30) attenuates the adverse effects of estrogen loss on cardiac fibrosis and diastolic dysfunction in mRen2.Lewis rats. This study was designed to address the effects of GPR30 on cardiac fibroblast proliferation in rats. The expression of GPR30 in cardiac fibroblasts isolated from adult Sprague–Dawley rats was confirmed by RT-PCR, Western blot analysis, and immunofluorescence staining. Results from BrdU incorporation assays, cell counting, carboxyfluorescein diacetate succinimidyl ester labeling in conjunction with flow cytometry, and Ki-67 staining showed that treatment with G1, a specific agonist of GPR30, inhibited cardiac fibroblast proliferation in a dose-dependent manner, which was associated with decreases in CDK1 and cyclin B1 protein expressions. In the GPR30-KO cells, BrdU incorporation, and CDK1 and cyclin B1 expressions significantly increased when compared to GPR30-intact cells. G1 had no effect on BrdU incorporation, CDK1 and cyclin B1 mRNA levels in GPR30-KO cells. In vivo studies showed increases in CDK1 and cyclin B1 mRNA levels, Ki-67-positive cells, and the immunohistochemistry staining of vimentin, a fibroblast marker, in the left ventricles from ovariectomized mRen2.Lewis rats versus hearts from ovary-intact littermates; 2 weeks of G1 treatment attenuated these adverse effects of estrogen loss. This study demonstrates that GPR30 is expressed in rat cardiac fibroblasts, and activation of GPR30 limits proliferation of these cells likely via suppression of the cell cycle proteins, cyclin B1, and CDK1.

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Acknowledgments

This work was funded in whole or part by National Institutes of Health Grants AG-042758 (to L. Groban), AG-033727 (to L. Groban), Doctoral Research Grant of Shandong Province BS2010YY005 (to Z. Zhao), and National Natural Science Foundation of China Grant 81270175 (to Z. Zhao). The funding sources had no involvement in the study design, in the collection, analysis, and interpretation of the data, nor in the writing of the report and decision to submit.

Conflict of interests

There are no conflicts of interest that could be perceived as prejudicing the impartiality of the research reported.

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Author notes

  1. Zhuo Zhao

    Present address: Department of Cardiology, Jinan Central Hospital, Affiliated with Shandong University, 105 Jiefang Road, Jinan, China

Authors and Affiliations

  1. Department of Anesthesiology, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC, 27157-1009, USA

    Hao Wang, Zhuo Zhao, Marina Lin & Leanne Groban

  2. Department of Internal Medicine-Molecular Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC, 27157-1009, USA

    Hao Wang & Leanne Groban

  3. The Hypertension and Vascular Research Center, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC, 27157, USA

    Leanne Groban

  4. Women’s Health Center of Excellence, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC, 27157, USA

    Leanne Groban

  5. The Sticht Center on Aging, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC, 27157, USA

    Leanne Groban

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  1. Hao Wang
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Correspondence to Leanne Groban.

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Wang, H., Zhao, Z., Lin, M. et al. Activation of GPR30 inhibits cardiac fibroblast proliferation. Mol Cell Biochem 405, 135–148 (2015). https://doi.org/10.1007/s11010-015-2405-3

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