MicroRNA-15a-5p induces pulmonary artery smooth muscle cell apoptosis in a pulmonary arterial hypertension model via the VEGF/p38/MMP-2 signaling pathway

摘要

The aim of the present study was to investigate the role of microRNA-15a-5p (miR-15a-5p) in pulmonary arterial hypertension (PAH) and elucidate the underlying pro-apoptotic mechanism. Reverse transcription-quantitative PCR analysis and gene microarray hybridization were used to measure the expression of miR-15a-5p in the lung tissues of rats with monocrotaline (MCT)-induced PAH. Flow cytometry and caspase-3/9 activity assays were adopted to measure the apoptosis of pulmonary artery smooth muscle cells (PASMCs). The expression of apoptosis-related proteins was analyzed using western blotting. The results demonstrated that the expression of miR-15a-5p was significantly increased in the lung tissues of rats with MCT-induced PAH. In addition, the overexpression of miR-15a-5p reduced PASMC proliferation, induced apoptosis, promoted the activity of caspase-3/9, induced the protein expression of B-cell lymphoma 2-associated X protein (Bax), decreased the expression of B-cell lymphoma 2 (Bcl-2), increased inflammation, as indicated by the expression of tumor necrosis factor-alpha (TNF)-alpha and interleukin (IL)-1 beta, IL-6 and IL-18, suppressed the protein expression of vascular endothelial growth factor (VEGF), and promoted the protein expression levels of phosphorylated (p)-p38 mitogen-activated protein kinase (p38) and matrix metalloproteinase (MMP)-2 in the PASMCs of rats with MCT-induced PAH. By contrast, the downregulation of miR-15a-5p increased cell proliferation, decreased apoptosis, reduced the activity of caspase-3/9 and the protein expression of Bax, increased the expression of Bcl-2, inhibited inflammation (as suggested by the expression of TNF-alpha, IL-1 beta, IL-6 and IL-18), induced the protein expression of VEGF, and suppressed the protein expression of p-p38 and MMP-2 in the PASMCs of rats with MCT-induced PAH. The inhibition of VEGF attenuated the effects of the overexpression of miR-15a-5p on the inhibition of cell proliferation, apoptotic rate, caspase-3/9 activity and protein expression of Bax, and it attenuated the increased inflammation, as indicated by the protein expression of p38 and MMP-2 in the PASMCs. In conclusion, the data of the present study demonstrated that miR-15a-5p induced the apoptosis of PASMCs in an animal model of PAH via the VEGF/p38/MMP-2 signaling pathway. However, further research is required to fully elucidate the role of miR-15a-5p in the development of PAH.