MicroRNA‐34a promotes CMECs apoptosis and upregulate inflammatory cytokines, thus worsening CMECs damage and inhibiting angiogenesis by negatively targeting the Notch signaling pathway

OBJECTIVE: Recently, microRNA-34a (miR-34a) has been reported to lead to secretion of proinflammatory cytokine in endothelial cells, whereas whether miR-34a plays a protective role in damaged cardiac microvascular endothelial cells (CMECs) remains to be determined. Herein, the purpose of this study is to explore the effect of miR-34a in mediating Notch signaling pathway in apoptosis and angiogenesis of damaged CMECs. METHODS: The primary mice CMECs were isolated, cultivated, and identified before establishment of damaged CMEC model by incubation with homocysteine (HCY) for 24 hours. Quantitative reverse-transcription polymerase chain reaction and Western blot analysis were applied to determine the expressions of miR-34a and Notch1. Cell viability and cell apoptosis were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and Hochest 33258 staining. Capillary-like structures formation assay was used to detect the capillary-like structures in CMECs. The expressions of inflammatory cytokines and angiogenesis factors were determined by enzyme-linked immunosorbent assay. RESULTS: In contrast to the blank group, the HCY and negative control groups demonstrated with elevated expressions of miR-34a, interleukin (IL)-1β, IL-6, and increased cell apoptosis rate, but decreased expressions of Notch1, IL-10, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and human growth factor (HGF), as well as attenuated cell viability and capillary-like structures of cells formation ability. In comparison with HCY group, the expressions of miR-34a, IL-1β, IL-6, and apoptosis rate were increased, whereas the expressions of Notch1, VEGF, bFGF, HGF, cell viability, and capillary-like structures of cells formation were inhibited in miR-34a mimic group. CONCLUSION: This study demonstrates that miR-34a can promote CMEC apoptosis and upregulate inflammatory cytokines, thus worsening CMEC damage and inhibiting angiogenesis by negatively targeting the Notch signaling pathway.