Moxibustion's protective role against atherosclerosis: Inhibition of Ca2+ overload‐triggered oxidative stress and inflammatory response via P2Y12/PI3K/AKT pathway

Abstract Objective This study aims to investigate the protective mechanism of moxibustion in combating atherosclerosis (AS). Methods Apolipoprotein E (ApoE)‐deficient mice, aged 8 weeks, were randomly assigned into four groups: the model group ( n = 6), SC79 group ( n = 6), moxibustion group ( n = 6), and moxibustion+SC79 group ( n = 6). All mice were fed with a high‐fat diet (HFD). Concurrently, 8‐week‐old C57BL/6 mice of the same genetic background were utilized as the control group ( n = 6) and were given a regular diet. Macrophages were isolated via flow cytometry. The intracellular Ca 2+ expression in macrophages was evaluated, and aortic plaques were quantitatively assessed through en face oil red O and Masson staining. The presence of macrophages and smooth muscle cells in AS plaques was determined by MAC‐3 and α‐smooth muscle actin (α‐SMA) immunohistochemistry. The relative fluorescence intensity of nuclear factor‐κB (NF‐κB) in macrophages was identified by immunofluorescence staining. The expressions of proteins related to the P2Y12/phosphatidylinositol 3‐hydroxy kinase (PI3K)/protein kinase B (AKT) signaling pathway were examined by Western blotting. Results Moxibustion reduced free Ca 2+ expression in macrophage cytoplasm, inhibiting Ca 2+ influx and oxidative stress. Significant reductions in atherosclerotic plaque formation and inflammation markers, including TNF‐α and IL‐1β, were noted in the moxibustion group. Moxibustion modulated the P2Y12/PI3K/AKT pathway, impacting various inflammatory and oxidative stress‐related proteins. Introduction of the AKT activator SC79 counteracted moxibustion's benefits, highlighting the P2Y12/PI3K/AKT pathway's central role. Conclusion Moxibustion, through the P2Y12/PI3K/AKT signaling pathway, can inhibit Ca 2+ overload‐induced oxidative stress and inflammatory response, decrease macrophage infiltration, and increase the content of smooth muscle cells and collagen, thereby exerting a protective effect against AS.