Dysregulation of miR-135a-5p promotes the development of rat pulmonary arterial hypertension in vivo and in vitro

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Abstract

Pulmonary arterial hypertension (PAH) is the most common form of pulmonary hypertension. Pulmonary arterial remodeling is closely related to the abnormal proliferation of pulmonary artery smooth muscle cells (PASMCs), which leads to the thickening of the medial layer of muscular arteries and then results in the narrowing or occlusion of the precapillary arterioles and PAH. However, the mechanisms underlying the abnormal proliferation of PASMCs remain unclear. In this study, we established rat primary PAH models using monocrotaline (MCT) injection or hypoxic exposure, then investigated the expression patterns of seven miRNAs associated with multiple pathogenic pathways central to pulmonary hypertension, and further explored the roles and the possible mechanisms of miR-135a during the development of PAH. In the rat primary PAH models, we observed that the expression of miR-135a-5p in lungs was drastically decreased at the initial stage of PAH development after MCT administration or hypoxic exposure, but it increased by 12-fold or 10-fold at the later stage. In vitro study in PASMCs showed a similar pattern of miR-135a-5p expression, with downregulation at 6 h but upregulation at 18, 24, and 48 h after hypoxic exposure. Early, but not late, administration of a miR-135a-5p mimic inhibited hypoxia-induced proliferation of PASMCs. The protective role of early miR-135a-5p agomir in the PAH rat model further supported the hypothesis that the early decrease in the expression of miR-135a-5p contributes to the proliferation of PASMCs and development of PAH, as early administration of miR-135a-5p agomir (10 nM, i.v.) reversed the elevated mean pulmonary arterial pressure and pulmonary vascular remodeling in MCT-treated rats. We revealed that miR-135a-5p directly bound to the 3′-UTR sequence of rat transient receptor potential channel 1 (TRPC1) mRNA and decreased TRPC1 protein expression, thus inhibiting PASMC proliferation. Collectively, our data suggest that dysregulation of miR-135a-5p in PASMCs contributes to the abnormal proliferation of PASMCs and the pathogenesis of PAH. Increasing miR-135a-5p expression at the early stage of PAH is a potential new avenue to prevent PAH development.

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Acknowledgements

This work was supported by the Chinese National Major New Drugs Innovation Projects [2011ZXJ09105X to Y. Tang], National Natural Science Foundation of China [81503083 to Y. Deng, 81372272 to J. Yan, and 81273507 to X. Li] and Scientific and Technological Research Program of Chongqing Municipal Education Commission [KJ1601703 to N. Liao]. Thanks for You-cai DENG's language editing.

Author contributions

H.-m.L. and Y.J. designed the research and conducted the experiments; Y.-x.Z. performed the infection assay; J.Y. and N.L. analyzed the data and wrote the paper; X.-h.L. and Y.T. designed the research, supervised the study, and wrote the manuscript.

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Correspondence to Xiao-hui Li.

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The authors declare no other commercial or financial conflict of interest.

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Keywords

  • pulmonary arterial hypertension
  • miR-135a-5p
  • pulmonary artery smooth muscle cells
  • transient receptor potential channel 1

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