弓形虫（T. gondii）是一种依赖寄生的细胞内原生动物寄生虫，可以引起肺部弓形虫病，其致病机制尚不完全了解。弓形虫病目前无法治愈。苦荞籽提取的植物多酚coixol具有多种生物活性，但其对T. gondii感染的影响尚未明确。本研究使用T. gondii RH菌株感染小鼠巨噬细胞系（RAW 264.7）和BALB/c小鼠，分别建立体外和体内感染模型，探讨coixol对T. gondii感染引起的肺损伤的保护作用和潜在机制。 本研究使用实时定量PCR、分子对接、局部表面等离子共振、共免疫沉淀、酶联免疫吸附测定、西方印迹和免疫荧光显微镜等方法，研究coixol的抗T. gondii效应和潜在的抗炎机制。结果表明，coixol抑制T. gondii负载和T.g.HSP70表达，减少炎症细胞的招募和浸润，并改善T. gondii感染引起的肺病理损伤。coixol可以直接结合T.g.HSP70或Toll样受体4（TLR4），从而破坏它们之间的相互作用。coixol通过抑制TLR4 /核因子（NF）-κB信号通路的激活，防止诱导型一氧化氮合酶、肿瘤坏死因子-α和高迁移率族-1的过表达，与TLR4抑制剂CLI-095的效果一致。这些结果表明，coixol通过干扰T.g.HSP70介导的TLR4 / NF-κB信号转导途径，改善T. gondii感染引起的肺损伤。总之，这些发现表明，coixol是治疗弓形虫病的一种有前途的有效前导化合物。版权所有©Elsevier B.V. 2023，保留所有权利。

Toxoplasma gondii (T. gondii) is an obligate intracellular protozoan parasite that causes pulmonary toxoplasmosis, although its pathogenesis is incompletely understood. There is no cure for toxoplasmosis. Coixol, a plant polyphenol extracted from coix seeds, has a variety of biological activities. However, the effects of coixol on T. gondii infection have not been clarified. In this study, we infected a mouse macrophage cell line (RAW 264.7) and BALB/c mice with the T. gondii RH strain to establish infection models in vitro and in vivo, respectively, to explore protective effects and potential mechanisms of coixol on lung injury caused by T. gondii infection. Anti-T. gondii effects and underlying anti-inflammatory mechanisms of coixol were investigated by real-time quantitative PCR, molecular docking, localized surface plasmon resonance, co-immunoprecipitation, enzyme-linked immunosorbent assay, western blotting, and immunofluorescence microscopy. The results show that coixol inhibits T. gondii loads and T. gondii-derived heat shock protein 70 (T.g.HSP70) expression. Moreover, coixol reduced inflammatory cell recruitment and infiltration, and ameliorated pathological lung injury induced by T. gondii infection. Coixol can directly bind T.g.HSP70 or Toll-like receptor 4 (TLR4) to disrupt their interaction. Coixol prevented overexpression of inducible nitric oxide synthase, tumor necrosis factor-α, and high mobility group box 1 by inhibiting activation of the TLR4/nuclear factor (NF)-κB signaling pathway, consistent with effects of the TLR4 inhibitor CLI-095. These results indicate that coixol improves T. gondii infection-induced lung injury by interfering with T.g.HSP70-mediated TLR4/NF-κB signaling. Altogether, these findings suggest that coixol is a promising effective lead compound for the treatment of toxoplasmosis.Copyright © 2023 Elsevier B.V. All rights reserved.