Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Medicine School of Shihezi University, Shihezi, Xinjiang 832008, P.R. China.
The 3rd Department of Cardiology, The First Affiliated Hospital of The Medical College, Shihezi University, Shihezi, Xinjiang 832008, P.R. China.
Mol Med Rep. 2020 May;21(5):2103-2112. doi: 10.3892/mmr.2020.11023. Epub 2020 Mar 12.
Angiotensin II (AngII) serves an important inflammatory role in cardiovascular disease; it can induce macrophages to differentiate into the M1‑type, produce inflammatory cytokines and resist pathogen invasion, and can cause a certain degree of damage to the body. Previous studies have reported that connexin 43 (Cx43) and NF‑κB (p65) are involved in the AngII‑induced inflammatory pathways of macrophages; however, the mechanisms underlying the effects of Cx43 and NF‑κB (p65) on AngII‑induced macrophage polarization have not been determined. Thus, the present study aimed to investigate the effects of Cx43 and NF‑κB (p65) on the polarization process of AngII‑induced macrophages. The macrophage polarization‑related proteins and mRNAs were examined by flow cytometry, western blotting, immunofluorescence, ELISA and reverse transcription‑quantitative PCR analyses. RAW264.7 macrophages were treated with AngII to simulate chronic inflammation and it was subsequently found that AngII promoted RAW 264.7 macrophage polarization towards the M1‑type by such effects as the release of inducible nitric oxide synthase (iNOS), tumour necrosis factor (TNF)‑α, IL‑1β, the secretion of IL‑6, and the expression of M1‑type indicators, such as CD86. Simultaneously, compared with the control group, the protein expression levels of Cx43 and phosphorylated (p)‑p65 were significantly increased following AngII treatment. The M1‑related phenotypic indicators, iNOS, TNF‑α, IL‑1β, IL‑6 and CD86, were inhibited by the NF‑κB (p65) signalling pathway inhibitor BAY117082. Similarly, the Cx43 inhibitors, Gap26 and Gap19, also inhibited the expression of M1‑related factors, and the protein expression levels of p‑p65 in the Gap26/Gap19 groups were significantly decreased compared with the AngII group. Altogether, these findings suggested that AngII may induce the polarization of RAW264.7 macrophages to the M1‑type through the Cx43/NF‑κB (p65) signalling pathway.
血管紧张素 II（AngII）在心血管疾病中发挥着重要的炎症作用；它可以诱导巨噬细胞分化为 M1 型，产生炎症细胞因子并抵抗病原体入侵，并且会对身体造成一定程度的损伤。先前的研究报道，连接蛋白 43（Cx43）和核因子-κB（p65）参与了巨噬细胞中 AngII 诱导的炎症途径；然而，Cx43 和 NF-κB（p65）对 AngII 诱导的巨噬细胞极化的影响的机制尚不清楚。因此，本研究旨在探讨 Cx43 和 NF-κB（p65）对 AngII 诱导的巨噬细胞极化过程的影响。通过流式细胞术、Western blot 印迹、免疫荧光、ELISA 和逆转录-定量 PCR 分析来检测巨噬细胞极化相关蛋白和 mRNA。用 AngII 处理 RAW264.7 巨噬细胞以模拟慢性炎症，结果发现 AngII 通过促进诱导型一氧化氮合酶（iNOS）、肿瘤坏死因子（TNF）-α、IL-1β 的释放、IL-6 的分泌以及 CD86 等 M1 型标志物的表达，促进 RAW264.7 巨噬细胞向 M1 型极化。同时，与对照组相比，AngII 处理后 Cx43 和磷酸化（p）-p65 的蛋白表达水平显著增加。M1 相关表型标志物，如 iNOS、TNF-α、IL-1β、IL-6 和 CD86，被 NF-κB（p65）信号通路抑制剂 BAY117082 抑制。同样，Cx43 抑制剂 Gap26 和 Gap19 也抑制了 M1 相关因子的表达，并且与 AngII 组相比，Gap26/Gap19 组中 p-p65 的蛋白表达水平显著降低。综上所述，这些发现表明 AngII 可能通过 Cx43/NF-κB（p65）信号通路诱导 RAW264.7 巨噬细胞向 M1 型极化。
