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MicroRNA‐34a promotes CMECs apoptosis and upregulate inflammatory cytokines, thus worsening CMECs damage and inhibiting angiogenesis by negatively targeting the Notch signaling pathway
Author(s) -
Li Jia,
Gong Jin,
Li Xiaobing,
Shen Li,
Xie Yewei,
Zhang Rufang
Publication year - 2019
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.27433
Subject(s) - viability assay , angiogenesis , proinflammatory cytokine , apoptosis , vascular endothelial growth factor , cytokine , microbiology and biotechnology , biology , endothelial stem cell , signal transduction , cell growth , western blot , notch signaling pathway , chemistry , immunology , cancer research , inflammation , biochemistry , in vitro , gene , vegf receptors
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.