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Estrogen administration reduces the risk of pulmonary arterial hypertension by modulating the miR-133a signaling pathways in rats

Gene Therapy volume 27, pages 113–126 (2020)Cite this article

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Abstract

We aimed to investigate how estrogen (ES) is implicated in the pathogenesis of pulmonary arterial hypertension (PAH) potentially by reducing the extent of vascular remodeling in females. HE assay, Western Blot, IHC, and real-time PCR were carried out to observe the role of ES in regulating miR-133a expression and the levels of MYOSLID, SRF, CTGF, and vascular remodeling in rats. In addition, MTT assay and flow cytometry were utilized to observe how ES affects cell proliferation and cell cycle in PAH. Moreover, luciferase assays were carried out to clarity the regulatory relationship between miR-133a and its downstream targets. ES administration relieved the deregulation of miR-133a, MYOSLID, SRF, and CTGF in PAH rats. In addition, ES also reduced the thickening of blood vessels in PAH rats. ES could activate miR-133a promoter and arrest the cells in the G0/G1 cycle, thus dose-dependently suppressing the proliferation of cells. In addition, the presence of ES, MYOSLID siRNA, or miR-133a precursor all altered the expression of MYOSLID, SP1, SRF, and CTGF, thus establishing a molecular signaling pathway among these factors. Furthermore, miR-133a could bind to SP1, MYOSLID, SRF, and CTGF to reduce their expression. Moreover, SRF was proved to function as an activator of miR-133a promoter. Two feedback loops were established in this study: a negative feedback loop between SRF and miR-133a, and a positive loop among miR-133a/SRF/MLK1/MYOSLID. ES treatment upregulates miR-133a expression and reduces the incidence of PAH and vascular remodeling.

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Funding

This research was supported by the fund from the Natural Science Foundation of China (No. 81300041, 81970051), the fund for Excellent top talent cultivation project of Anhui Higher Education Institutions (gxyqZD2017030).

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  1. These authors contributed equally: Sijing Zhou, Ke Zhu

Authors and Affiliations

  1. Hefei Prevention and Treatment Center for Occupational Diseases, 230022, Hefei, China

    Sijing Zhou

  2. Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Anhui Medical University, 230022, Hefei, China

    Ke Zhu, Huihui Jiang, Peipei Wu, Xing Ding, Li Sun, Gengyun Sun & Ran Wang

  3. Department of General Medicine, Hefei Second People’s Hospital, Changjiang East Road, 230022, Hefei, China

    Yongsheng Du & Aiqun Xu

  4. Department of Oncology, The First Affiliated Hospital of Anhui Medical University, 230022, Hefei, China

    Min Li

  5. Department of Respiratory Medicine, Ningbo First Hospital, 315000, Ningbo, China

    Chao Cao

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Zhou, S., Zhu, K., Du, Y. et al. Estrogen administration reduces the risk of pulmonary arterial hypertension by modulating the miR-133a signaling pathways in rats. Gene Ther 27, 113–126 (2020). https://doi.org/10.1038/s41434-019-0103-6

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