Deng Liangyong, Yu Qiulei, Kuang Gang, Wang Liuyang, Fan Jing, Ye Lin
Department of Critical Care Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
Department of Pathology, Jinshan Hospital, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
Inflamm Res. 2025 Mar 28;74(1):59. doi: 10.1007/s00011-025-02026-3.
Luteolin has an anti-inflammatory effect, but the mechanism has not been elucidated in sepsis-induced acute hepatic injury (AHI). The purpose of this study was to investigate the effects and potential mechanisms of luteolin on sepsis-induced AHI.
In this study, we utilized both wild-type (WT) mice and Toll-like receptor 4 (TLR4)-deficient (TLR4) mice alongside RAW264.7 cells. We constructed a CLP-induced AHI mouse model to study the effects of luteolin on liver inflammation, survival and liver macrophage subtypes in mice. In addition, we extracted mouse serum, mouse bone marrow-derived macrophages (BMDMs) and liver tissue and analysed the effects of luteolin on macrophage polarization subtypes and downstream inflammatory cytokines by flow cytometry, ELISA, Western blotting (WB) and qPCR. To further verify the effect of luteolin on macrophage polarization and explore the possible potential mechanism, we used a CLP-induced AHI mouse model and LPS-stimulated RAW 264.7 macrophages to assess the effect of luteolin on macrophage polarization; the expression of TNF-α and IL-10 in the cell culture supernatant; and the expression of iNOS, ARG-1, NF-κB (P65), p-P65 and MyD88 by flow cytometry, ELISA, immunohistochemistry and Western blotting.
We found that luteolin reduced liver injury and inflammatory response and improved the survival rate of mice. Luteolin modulated the macrophage subtype proportion, promoted the change of macrophages from a proinflammatory M1 phenotype to an anti-inflammatory M2 phenotype, and reduced the inflammatory response both in vivo and in vitro. Moreover, luteolin reduced the expression of NF-κB (p-P65), TLR4 and MyD88. By integrating the predictions from network pharmacology with the in vitro and in vivo experimental results, it was determined that the mechanism by which luteolin alleviates sepsis-induced acute hepatic injury is closely related to the TLR4/MyD88/NF-κB pathway.
The results of this study suggest that luteolin helps alleviate liver injury, reduces the expression of proinflammatory cytokines and promotes the expression of anti-inflammatory factors in sepsis-induced acute hepatic injury. This effect may be related to the regulation of macrophage polarization by luteolin through the TLR4/MyD88/NF-κB signalling pathway.
木犀草素具有抗炎作用，但脓毒症诱导的急性肝损伤（AHI）中的作用机制尚未阐明。本研究旨在探讨木犀草素对脓毒症诱导的AHI的影响及潜在机制。
在本研究中，我们使用野生型（WT）小鼠、Toll样受体4（TLR4）缺陷型（TLR4-/-）小鼠以及RAW264.7细胞。构建盲肠结扎穿孔（CLP）诱导的AHI小鼠模型，以研究木犀草素对小鼠肝脏炎症、存活率及肝脏巨噬细胞亚型的影响。此外，提取小鼠血清、小鼠骨髓来源的巨噬细胞（BMDMs）及肝脏组织，通过流式细胞术、酶联免疫吸附测定（ELISA）、蛋白质免疫印迹法（WB）及定量聚合酶链反应（qPCR）分析木犀草素对巨噬细胞极化亚型及下游炎症细胞因子的影响。为进一步验证木犀草素对巨噬细胞极化的作用并探索可能的潜在机制，我们使用CLP诱导的AHI小鼠模型及脂多糖（LPS）刺激的RAW 264.7巨噬细胞来评估木犀草素对巨噬细胞极化的影响；细胞培养上清液中肿瘤坏死因子-α（TNF-α）和白细胞介素-10（IL-10）的表达；以及通过流式细胞术、ELISA、免疫组织化学和WB检测诱导型一氧化氮合酶（iNOS）、精氨酸酶-1（ARG-1）、核因子κB（NF-κB，P65）、磷酸化核因子κB（p-P65）和髓样分化因子88（MyD88）的表达。
我们发现木犀草素减轻了肝脏损伤和炎症反应，并提高了小鼠的存活率。木犀草素调节巨噬细胞亚型比例，促进巨噬细胞从促炎M1表型向抗炎M2表型转变，并在体内和体外均减轻了炎症反应。此外，木犀草素降低了NF-κB（p-P65）、TLR4和MyD88的表达。通过将网络药理学预测结果与体外和体内实验结果相结合，确定木犀草素减轻脓毒症诱导的急性肝损伤的机制与TLR4/MyD88/NF-κB通路密切相关。
本研究结果表明，木犀草素有助于减轻脓毒症诱导的急性肝损伤中的肝损伤，降低促炎细胞因子的表达，并促进抗炎因子的表达。这种作用可能与木犀草素通过TLR4/MyD
