Angiotensin II induces RAW264.7 macrophage polarization to the M1-type through the connexin 43/NF-κB pathway

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.