失调的炎症在许多疾病的发病机制中是很重要的，包括癌症、过敏和自身免疫病。巨噬细胞的激活和极化通常涉及炎症的启动、维持和解决。Perhexiline (PHX)是一种抗心绞痛药物，被认为可以调节巨噬细胞功能，但PHX对巨噬细胞的分子影响尚不清楚。在本研究中，我们研究了PHX治疗对巨噬细胞激活和极化的影响，并揭示了PHX引起的潜在蛋白质组变化。我们使用已建立的方案，将人类THP-1单核细胞分化为M1或M2型巨噬细胞，包括三个不同的、连续的阶段(预处理、休息和分化)。我们使用流式细胞术、定量聚合酶链反应(qPCR)和酶联免疫吸附试验(ELISA)研究了PHX治疗对M1或M2型巨噬细胞极化的影响。使用数据独立采集质谱(DIA MS)研究蛋白质的定量变化。PHX治疗促进了M1型巨噬细胞的极化，包括增加STAT1和CCL2的表达和IL-1β的分泌。当PHX添加到M1培养的分化阶段时，才出现这种效应。PHX处理的M1型培养物的蛋白质组分析显示代谢途径(脂肪酸代谢、胆固醇稳态和氧化磷酸化)和免疫信号通路(受体酪氨酸激酶、Rho GTP酰化酶和干扰素)发生了变化。这是第一项报道PHX对THP-1巨噬细胞极化作用以及这些细胞蛋白质组变化的研究。版权所有 © 2023 Dhakal，Li，Ramezanpour，Houtak，Li，Bouras，Collela，Chegeni，Chataway，Drew，Sallustio，Vreugde，Smith，Maddern，Licari和Fenix。

Dysregulated inflammation is important in the pathogenesis of many diseases including cancer, allergy, and autoimmunity. Macrophage activation and polarisation are commonly involved in the initiation, maintenance and resolution of inflammation. Perhexiline (PHX), an antianginal drug, has been suggested to modulate macrophage function, but the molecular effects of PHX on macrophages are unknown. In this study we investigated the effect of PHX treatment on macrophage activation and polarization and reveal the underlying proteomic changes induced.We used an established protocol to differentiate human THP-1 monocytes into M1 or M2 macrophages involving three distinct, sequential stages (priming, rest, and differentiation). We examined the effect of PHX treatment at each stage on the polarization into either M1 or M2 macrophages using flow cytometry, quantitative polymerase chain reaction (qPCR) and enzyme linked immunosorbent assay (ELISA). Quantitative changes in the proteome were investigated using data independent acquisition mass spectrometry (DIA MS).PHX treatment promoted M1 macrophage polarization, including increased STAT1 and CCL2 expression and IL-1β secretion. This effect occurred when PHX was added at the differentiation stage of the M1 cultures. Proteomic profiling of PHX treated M1 cultures identified changes in metabolic (fatty acid metabolism, cholesterol homeostasis and oxidative phosphorylation) and immune signalling (Receptor Tyrosine Kinase, Rho GTPase and interferon) pathways.This is the first study to report on the action of PHX on THP-1 macrophage polarization and the associated changes in the proteome of these cells.Copyright © 2023 Dhakal, Li, Ramezanpour, Houtak, Li, Bouras, Collela, Chegeni, Chataway, Drew, Sallustio, Vreugde, Smith, Maddern, Licari and Fenix.