阿匹莫德通过双重抑制PI3K/AKT通路及整合素信号缓解ApoE小鼠动脉粥样硬化

Apilimod Attenuates Atherosclerosis in ApoE-Deficient Mice via Dual Inhibition of the PI3K/AKT Pathway and Integrin Signaling?

关键词:  阿匹莫德  动脉粥样硬化  PI3K/AKT通路  炎症反应  血管平滑肌细胞增殖

Key words:  Apilimod  Atherosclerosis  PI3K/AKT pathway  Inflammatory response  Vascular smooth muscle cell proliferation

摘要:

背景与目的：动脉粥样硬化（Atherosclerosis，AS）的病理进程涉及高脂血症、慢性炎症及血管平滑肌细胞（Vascular smooth muscle cells，VSMC）异常增殖。阿匹莫德（Apilimod）作为IL-12/IL-23与PIKfyve的双重抑制剂，其抗AS机制尚不明确。本研究旨在揭示阿匹莫德调控AS的潜在靶点及作用通路。 方法：通过SuperPred数据库预测阿匹莫德靶标基因，结合KEGG富集分析与动脉粥样硬化基因集筛选核心靶点。采用MOE分子对接与表面等离子共振技术（Surface plasmon resonance，SPR）检测阿匹莫德与PI3K/AKT通路成员的相互作用。通过氧化修饰低密度脂蛋白（ox-LDL）诱导的VSMC增殖模型、急性高脂血小鼠及ApoE-/-小鼠模型，评估阿匹莫德对脂质代谢、炎症因子及细胞周期的影响。 结果：阿匹莫德靶标显著富集于PI3K/AKT通路，分子对接显示其与ITGAV、PIK3CA、AKT1、CDK4等靶点高亲和力结合（结合能＜-5 kcal/mol，RMSD＜2 ?）；SPR结果验证阿匹莫德与PI3K，AKT1以及ITGAV蛋白之间存在高亲和相互作用。阿匹莫德显著抑制ox-LDL诱导的VSMC增殖（P<0.01），阻滞G1/S期转换（P<0.05）。在ApoE-/-小鼠中，阿匹莫德显著降低血清IL-6、TNF-α及主动脉ICAM1/VCAM1表达（P<0.05/P<0.01），减少主动脉脂质沉积（降幅31%-35%）。阿匹莫德下调主动脉中CDK4、CCND1、EGFR的转录与蛋白表达（P<0.05），抑制PI3K/AKT-NF-κB/STAT3信号轴。 结论：阿匹莫德通过靶向PI3K/AKT通路及整合素家族，抑制炎症反应与VSMC增殖，缓解AS进展，但其作用独立于降脂机制。本研究为AS的靶向治疗提供了新策略，未来需优化其组织特异性递送并探索联合用药潜力。

Abstract:

BACKGROUND and OBJECTIVE: The pathogenesis of atherosclerosis (AS) involves hyperlipidemia, chronic inflammation, and abnormal proliferation of vascular smooth muscle cells (VSMCs). Apilimod, a dual inhibitor of IL-12/IL-23 and PIKfyve, has unclear mechanisms in combating AS. This study aimed to elucidate the potential targets and pathways by which apilimod regulates AS. METHODS: Target genes of apilimod were predicted using the SuperPred database, and core targets were screened through KEGG enrichment analysis and atherosclerosis-related gene sets. MOE molecular docking and surface plasmon resonance (SPR) validated interactions between apilimod and PI3K/AKT pathway components. The effects of apilimod on lipid metabolism, inflammatory cytokines, and cell cycle progression were assessed using oxidized low-density lipoprotein (ox-LDL)-induced VSMC proliferation, acute hyperlipidemic mouse models, and ApoE-deficient mice. RESULTS: Apilimod targets were significantly enriched in the PI3K/AKT pathway. Molecular docking revealed high-affinity binding (binding energy<-5kcal/mol, RMSD<2?) between apilimod and key targets, including ITGAV, PIK3CA, AKT1, and CDK4. The SPR results verified that there were highly affinity interactions between apilimod and PI3K, AKT1, and ITGAV proteins. Apilimod markedly inhibited ox-LDL-induced VSMC proliferation (P<0.01) and blocked the G1/S phase transition (P<0.05). In ApoE-deficient mice, Apilimod significantly reduced serum IL-6, TNF-α, and aortic expression of ICAM1/VCAM1 (P<0.05/P<0.01), with a 31%–35% decrease in aortic lipid deposition. Additionally, apilimod downregulated the transcription and protein expression of CDK4, CCND1, and EGFR (P < 0.05) and suppressed the PI3K/AKT-NF-κB/STAT3 signaling axis. CONCLUSION: Apilimod alleviates AS progression by targeting the PI3K/AKT pathway and integrin signaling, thereby inhibiting inflammation and VSMC proliferation. However, its efficacy is independent of lipid-lowering effects. This study provides a novel therapeutic strategy for AS, highlighting the need to optimize tissue-specific delivery and explore combination therapies in future research.