Liu Qiang, Liu Anli, Leng Shaoqiu, Zhang Xiaoyu, Wang Xiaolin, Cheng Zhang, Wang Shuwen, Peng Jun, Feng Qi
Department of Hematology, Qilu Hospital of Shandong University, Jinan, Shandong 250012,China.
Department of Hematology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong 250021, China.
Chin Med J (Engl). 2025 May 29. doi: 10.1097/CM9.0000000000003586.
Macrophage polarization anomalies and dysfunction play a crucial role in the pathogenesis of immune thrombocytopenia (ITP). Itaconate is a Krebs cycle-derived immunometabolite synthesized by myeloid cells to modulate cellular metabolism and inflammatory responses. This study aimed to evaluate the immunoregulatory effects of an itaconate derivative on macrophages in patients with ITP.
Peripheral blood-derived macrophages from patients with ITP and healthy controls were treated with 4-octyl itaconate (4-OI), a derivative of itaconate that can penetrate the cell membrane. Macrophage polarization, antigen-presenting functions, and phagocytic capability were measured via flow cytometry and enzyme-linked immunosorbent assay (ELISA). Macrophage glycolysis in patients with ITP and the metabolic regulatory effect of 4-OI were detected using a Seahorse XFe96 Analyzer. An active murine model of ITP was used to evaluate the therapeutic effects of 4-OI in vivo.
4-OI reduced the levels of CD80 and CD86 in M1 macrophages and suppressed the release of tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 pro-inflammatory cytokines, suggesting that 4-OI could hinder the polarization of macrophages toward an M1 phenotype. We found that 4-OI pretreated M1 macrophages reduced the proliferation of CD4+ T cells and promoted the differentiation of regulatory T cells. In addition, after 4-OI treatment, the phagocytic capacity of M1 macrophages toward antibody-coated platelets decreased significantly in patients with ITP. In addition, the glycolytic function of M1 macrophages was elevated in individuals with ITP compared to those in healthy controls. 4-OI treatment downregulated glycolysis in M1 macrophages. The glycolysis inhibitor 2-deoxy-d-glucose (2-DG) also inhibited the polarization of M1 macrophages and restored their functions. In vivo, 4-OI treatment significantly increased platelet counts in the active ITP murine model.
Itaconate derivative 4-OI inhibited M1 macrophage polarization and restored impaired functions through metabolic reprogramming. This study provides a novel therapeutic option for ITP.
巨噬细胞极化异常和功能障碍在免疫性血小板减少症（ITP）的发病机制中起关键作用。衣康酸是一种由三羧酸循环衍生的免疫代谢物，由髓样细胞合成，用于调节细胞代谢和炎症反应。本研究旨在评估衣康酸衍生物对ITP患者巨噬细胞的免疫调节作用。
用衣康酸衍生物4-辛基衣康酸（4-OI）处理ITP患者和健康对照者外周血来源的巨噬细胞，4-OI可穿透细胞膜。通过流式细胞术和酶联免疫吸附测定（ELISA）检测巨噬细胞极化、抗原呈递功能和吞噬能力。使用海马XFe96分析仪检测ITP患者巨噬细胞的糖酵解以及4-OI的代谢调节作用。采用ITP活性小鼠模型评估4-OI的体内治疗效果。
4-OI降低了M1巨噬细胞中CD80和CD86的水平，并抑制了肿瘤坏死因子-α（TNF-α）和白细胞介素（IL）-6促炎细胞因子的释放，表明4-OI可阻碍巨噬细胞向M1表型极化。我们发现，4-OI预处理的M1巨噬细胞可降低CD4+T细胞的增殖并促进调节性T细胞的分化。此外，4-OI处理后，ITP患者M1巨噬细胞对抗体包被血小板的吞噬能力显著降低。此外，与健康对照者相比，ITP患者M1巨噬细胞的糖酵解功能升高。4-OI处理下调了M1巨噬细胞的糖酵解。糖酵解抑制剂2-脱氧-D-葡萄糖（2-DG）也抑制了M1巨噬细胞的极化并恢复了其功能。在体内，4-OI治疗显著提高了活性ITP小鼠模型的血小板计数。
衣康酸衍生物4-OI通过代谢重编程抑制M1巨噬细胞极化并恢复受损功能。本研究为ITP提供了一种新的治疗选择。
