Xiong Xiaofang, Huang Wei, Yang Xuexue, Wang Xun, Wu Beibei, Li Dongsheng
The Department of Cardiology at Wuhan Third Hospital, Wuhan, Hubei, China.
The Department of Traditional Chinese Medicine at Wuhan Third Hospital, Wuhan, Hubei, China.
Front Pharmacol. 2025 Jul 10;16:1607349. doi: 10.3389/fphar.2025.1607349. eCollection 2025.
T cells are contributors to atherosclerosis pathogenesis. Granulocyte-macrophage-colony-stimulating factor (GM-CSF)-producing T helper (ThGM) cells, a specialized helper T cell subset that highly expresses GM-CSF but lacks other helper T cell markers, could exacerbate atherosclerosis development. Calycosin has been reported to suppress atherosclerosis progression. However, the effect of calycosin on ThGM cells is unknown. This study was designed to test the calycosin-induced impact on the pro-atherosclerotic function of ThGM cells in a mouse atherosclerosis model.
Apolipoprotein E knockout (ApoE) mice were fed a high-fat diet and calycosin. The phenotype and cytokine expression of aortic ThGM cells were assessed by flow cytometry. Calycosin-derived influences on ThGM cell differentiation, proliferation, and function were determined by flow cytometry, quantitative RT-PCR, Immunoblotting, gene silencing assays, and co-culture with macrophages.
Aortic ThGM cell frequency was attenuated after calycosin administration. Live aortic ThGM cells, phenotypically featuring CD4CCR6CCR8CXCR3CCR10, showed slower proliferation and weaker macrophage-activating capability in calycosin-treated mice. Besides, calycosin repressed ThGM cell differentiation and subsequently impaired ThGM cell-mediated macrophage activation, oxidized low-density lipoprotein (Ox-LDL) uptake, and foam cell formation. Importantly, calycosin upregulated nuclear receptor subfamily 4 group A member 3 (NR4A3) in ThGM cells. NR4A3 silencing partially restored the function of calycosin-treated ThGM cells.
Calycosin inhibits ThGM cell activity to suppress ThGM-cell-mediated activation of pro-atherosclerotic macrophages to ultimately ameliorate atherosclerosis progression. Therefore, we revealed a novel mechanism by which calycosin protects against atherosclerosis.
T细胞在动脉粥样硬化发病机制中起作用。产生粒细胞-巨噬细胞集落刺激因子（GM-CSF）的辅助性T（ThGM）细胞是一种特殊的辅助性T细胞亚群，其高表达GM-CSF但缺乏其他辅助性T细胞标志物，可加剧动脉粥样硬化的发展。据报道，毛蕊异黄酮可抑制动脉粥样硬化进展。然而，毛蕊异黄酮对ThGM细胞的作用尚不清楚。本研究旨在测试毛蕊异黄酮对小鼠动脉粥样硬化模型中ThGM细胞促动脉粥样硬化功能的影响。
给载脂蛋白E基因敲除（ApoE）小鼠喂食高脂饮食并给予毛蕊异黄酮。通过流式细胞术评估主动脉ThGM细胞的表型和细胞因子表达。通过流式细胞术、定量RT-PCR、免疫印迹、基因沉默试验以及与巨噬细胞共培养，确定毛蕊异黄酮对ThGM细胞分化、增殖和功能的影响。
给予毛蕊异黄酮后，主动脉ThGM细胞频率降低。在给予毛蕊异黄酮的小鼠中，具有CD4CCR6CCR8CXCR3CCR10表型特征的存活主动脉ThGM细胞增殖较慢，巨噬细胞激活能力较弱。此外，毛蕊异黄酮抑制ThGM细胞分化，随后损害ThGM细胞介导的巨噬细胞激活、氧化型低密度脂蛋白（Ox-LDL）摄取和泡沫细胞形成。重要的是，毛蕊异黄酮上调ThGM细胞中的核受体亚家族4A组成员3（NR4A3）。NR4A3沉默部分恢复了经毛蕊异黄酮处理的ThGM细胞的功能。
毛蕊异黄酮抑制ThGM细胞活性，以抑制ThGM细胞介导的促动脉粥样硬化巨噬细胞激活，最终改善动脉粥样硬化进展。因此，我们揭示了毛蕊异黄酮预防动脉粥样硬化的新机制。
