Department of Geratology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Biomed Res Int. 2020 Apr 1;2020:8718321. doi: 10.1155/2020/8718321. eCollection 2020.
Inflammatory responses play important roles in the pathogenesis of atherosclerosis. The purpose of this study was to investigate the relationship between microRNA-146b-3p (miR-146b-3p) and inflammatory factors in thrombosis.
THP-1 cells were cultured in vitro, Western blot was used to determine the protein levels of COX-2 and p38MAPK in the cells, and real-time PCR was used to detect the mRNA expression of miRNA-146b-3p and COX-2. A lentiviral expression vector of miRNA-146b-3p and its inhibitor were constructed to transfect THP-1 cells. COX-2 and p38MAPK expression in transfected cells was detected by Western blot and real-time PCR, respectively.
Ang II and TNF- could elevate the expression of COX-2 in monocytes. The expression of COX-2 was upregulated by p38MAPK, which could be phosphorylated by Ang II, while there was an increasing tendency of p38MAPK phosphorylation after TNF- stimulation. In addition, COX-2 expression and P38MAPK phosphorylation could be downregulated by miRNA-146b-3p and upregulated by the miRNA-146b-3p inhibitor. Ang II could increase miR-146b-3p expression, although there was no significant difference; however, the expression of miR-146b-3p was enhanced significantly by TNF-.
Our data implied that altered expression of miR-146b-3p was closely related to the progression of inflammation mediating the P38MAPK/COX-2 pathway. We suggest that the miR-146b-3p/p38MAPK/COX-2 pathway plays a key role in inflammation and arterial thrombosis.
炎症反应在动脉粥样硬化的发病机制中起着重要作用。本研究旨在探讨 microRNA-146b-3p（miR-146b-3p）与血栓形成中的炎症因子之间的关系。
体外培养 THP-1 细胞，Western blot 法测定细胞中 COX-2 和 p38MAPK 的蛋白水平，实时 PCR 法检测 miR-146b-3p 和 COX-2 的 mRNA 表达。构建 miR-146b-3p 的慢病毒表达载体及其抑制剂，转染 THP-1 细胞。Western blot 和实时 PCR 分别检测转染细胞中 COX-2 和 p38MAPK 的表达。
Ang II 和 TNF-可上调单核细胞中 COX-2 的表达。COX-2 的表达受 p38MAPK 上调，p38MAPK 可被 Ang II 磷酸化，而 TNF-刺激后 p38MAPK 磷酸化呈上升趋势。此外，COX-2 表达和 P38MAPK 磷酸化可被 miR-146b-3p 下调，被 miR-146b-3p 抑制剂上调。Ang II 可增加 miR-146b-3p 的表达，尽管差异无统计学意义；然而，TNF-可显著增强 miR-146b-3p 的表达。
我们的数据表明，miR-146b-3p 的改变表达与炎症进展密切相关，调节 P38MAPK/COX-2 通路。我们提出，miR-146b-3p/p38MAPK/COX-2 通路在炎症和动脉血栓形成中起着关键作用。
