疟疾相关的急性肺损伤/急性呼吸窘迫综合征（MA-ALI/ARDS）是恶性疟原虫感染的致命并发症，部分由巨噬细胞募集和极化引发。据报道，铜暴露会增加疟疾感染的风险，而铜积累引起的铜凋亡会引发 M1 巨噬细胞极化。因此推测铜凋亡可能是 MA-ALI/ARDS 发病机制中的关键介质，但其潜在机制仍不清楚。本研究旨在探讨铜凋亡在小鼠 MA-ALI/ARDS 严重程度中的作用。我们利用感染伯氏疟原虫 ANKA 的雌性 C57BL/6 小鼠建立 MA-ALI/ARDS 实验模型，并用有效的铜离子载体双硫仑 (DSF) 或铜离子螯合剂四硫代钼酸盐 (TTM)。在体外用受感染的红细胞 (iRBC) 刺激的 RAW 264.7 巨噬细胞也用 DSF-CuCl2 或 TTM-CuCl2 进行靶向，以进一步研究其潜在机制。我们的研究结果表明，铜的量和在实验性 MA-ALI/ARDS 小鼠的肺组织中，SLC31A1（铜流入转运蛋白）和 FDX1（铜凋亡的关键正调节因子）的表达显着降低，但 ATP7A（铜流出转运蛋白）的表达显着减少。与感染伯氏疟原虫 ANKA 的对照组相比，给予 DSF 的小鼠表现出寄生虫血症/肺寄生虫负荷、支气管肺泡灌洗液 (BALF) 中的总蛋白浓度、肺湿/干重比、血管渗漏和肺组织的病理变化。引人注目的是，接受 DSF 治疗的实验性 MA-ALI/ARDS 小鼠还表现出铜水平、SLC31A1 和 FDX1 表达、CD86、CD68、SLC31A1-CD68 和 FDX1-CD68 巨噬细胞数量以及信使 RNA (mRNA) 水平显着升高。肺组织中的促炎细胞因子（肿瘤坏死因子 [TNF-α] 和诱导型一氧化氮合酶 [iNOS]），但体重、存活时间、ATP7A 表达、CD206 巨噬细胞数量和 mRNA 水平显着下降抗炎细胞因子（转化生长因子 β [TGF-β] 和白细胞介素 10 [IL-10]）。相比之下，TTM 治疗逆转了受感染小鼠的这些变化。同样，体外实验显示，在 DSF-CuCl2 靶向的 iRBC 刺激的 RAW 264.7 细胞中，SLC31A1、FDX1、CD86、TNF-α 和 iNOS 的 mRNA 水平显着升高，但引发了 mRNA 水平的显着下降。 ATP7A、CD206、TGF-β 和 IL-10。相比之下，TTM-CuCl2 治疗也逆转了 iRBC 刺激的 RAW 264.7 细胞中的这些趋势。我们的数据表明，用 DSF 激活铜凋亡会部分诱导肺巨噬细胞的 M1 极化，从而加重 MA-ALI/ARDS 的严重程度，而抑制相反，TTM 治疗铜凋亡可通过促进巨噬细胞 M2 极化来改善 MA-ALI/ARDS 的严重程度。我们的研究结果表明，阻断铜凋亡可能是治疗 MA-ALI/ARDS 的潜在治疗策略。© 2024。作者。

Malaria-associated acute lung injury/acute respiratory distress syndrome (MA-ALI/ARDS) is a fatal complication of Plasmodium falciparum infection that is partially triggered by macrophage recruitment and polarization. As reported, copper exposure increases the risk of malaria infection, and copper accumulation-induced cuproptosis triggers M1 macrophage polarization. It is thus hypothesized that cuproptosis could act as a critical mediator in the pathogenesis of MA-ALI/ARDS, but its underlying mechanism remains unclear. The present study aimed to explore the role of cuproptosis in the severity of murine MA-ALI/ARDS.We utilized an experimental model of MA-ALI/ARDS using female C57BL/6 mice with P. berghei ANKA infection, and treated these animals with the potent copper ion carrier disulfiram (DSF) or copper ion chelator tetrathiomolybdate (TTM). The RAW 264.7 macrophages, which were stimulated with infected red blood cells (iRBCs) in vitro, were also targeted with DSF-CuCl2 or TTM-CuCl2 to further investigate the underlying mechanism.Our findings showed a dramatic elevation in the amount of copper and the expression of SLC31A1 (a copper influx transporter) and FDX1 (a key positive regulator of cuproptosis) but displayed a notable reduction in the expression of ATP7A (a copper efflux transporter) in the lung tissue of experimental MA-ALI/ARDS mice. Compared to the P. berghei ANKA-infected control group, mice that were administered DSF exhibited a remarkable increase in parasitemia/lung parasite burden, total protein concentrations in bronchoalveolar lavage fluid (BALF), lung wet/dry weight ratio, vascular leakage, and pathological changes in lung tissue. Strikingly, the experimental MA-ALI/ARDS mice with DSF treatment also demonstrated dramatically elevated copper levels, expression of SLC31A1 and FDX1, numbers of CD86+, CD68+, SLC31A1+-CD68+, and FDX1+-CD68+ macrophages, and messenger RNA (mRNA) levels of pro-inflammatory cytokines (tumor necrosis factor [TNF-α] and inducible nitric oxide synthase [iNOS]) in lung tissue, but showed a remarkable decrease in body weight, survival time, expression of ATP7A, number of CD206+ macrophages, and mRNA levels of anti-inflammatory cytokines (transforming growth factor beta [TGF-β] and interleukin 10 [IL-10]). In contrast, TTM treatment reversed these changes in the infected mice. Similarly, the in vitro experiment showed a notable elevation in the mRNA levels of SLC31A1, FDX1, CD86, TNF-α, and iNOS in iRBC-stimulated RAW 264.7 cells targeted with DSF-CuCl2, but triggered a remarkable decline in the mRNA levels of ATP7A, CD206, TGF-β, and IL-10. In contrast, TTM-CuCl2 treatment also reversed these trends in the iRBC-stimulated RAW 264.7 cells.Our data demonstrate that the activation of cuproptosis with DSF aggravated the severity of MA-ALI/ARDS by partially inducing M1 polarization of pulmonary macrophages, while inhibition of cuproptosis with TTM contrarily ameliorated the severity of MA-ALI/ARDS by promoting macrophage M2 polarization. Our findings suggest that blockage of cuproptosis could be a potential therapeutic strategy for treatment of MA-ALI/ARDS.© 2024. The Author(s).