Role of miR-199b-5p in regulating angiogenesis in mouse myocardial microvascular endothelial cells through HSF1/VEGF pathway

Our study explored effects of miR-199b-5p on angiogenesis in mouse myocardial microvascular endothelial cells (MMVECs) and the involved working mechanisms. We applied explant culture to incubate C57/BL6 mouse MMVECs. Lipofection was used to transfect miR-199b-5p mimic, miR-199b-5p inhibitor and miR-199b-5p scramble respectively. MMVECs were divided into miR-199b-5p up-regulation, miR-199b-5p down-regulation and control groups based on above sequence. Expressions of miR-199b-5p, heat shock factor protein 1 (HSF1) mRNA were assessed by real-time quantitative polymerase chain reaction (RT-QFCR). Expressions of HSF1 and vascular endothelial growth factor (VEGF) were assessed by Western Blotting. Cell proliferation was assessed by CCK8. Tubule formation assay was conducted to assess formation of blood vessels. Results showed that miR-199b-5p up/down-regulation groups exhibited no obvious differences in the expressions of HSF1 mRNA compared to control group. However, miR-199b-5p up-regulation group recorded lower expressions of HSF1 and VEGF in the level of protein, and reduced cell proliferation and tubule formation. Whereas, miR-199b-5p down-regulation group presented the contrary results. The experiment indicated that miR-199b-5p can regulate proliferation and angiogenesis in mouse MMVECs through the pathway of HSF1/VEGF. (C) 2016 Elsevier B.V. All rights reserved.