南方医科大学学报2025，Vol.45Issue(4)：692-701,10.DOI:10.12122/j.issn.1673-4254.2025.04.03
穗花杉双黄酮通过抑制细胞焦亡减轻小鼠急性肺损伤
Amentoflavone alleviates acute lung injury in mice by inhibiting cell pyroptosis
摘要
Abstract
Objective To investigate the effect of amentoflavone(AF)for alleviating lipopolysaccharide(LPS)-induced acute lung injury(ALI)in mice and inhibiting NLRP3/ASC/Caspase-1 axis-mediated pyroptosis.Methods Female BALB/c mice were randomly divided into control group,LPS group,and AF treatment groups at low,moderate and high doses(n=12).ALI models were established by tracheal LPS instillation,and in AF treatment groups,AF was administered by gavage 30 min before LPS instillation.Six hours after LPS instillation,the mice were euthanized for examining lung tissue histopathological changes,protein levels in BALF,and MPO levels in the lung tissue.In the in vitro experiment,RAW264.7 cells were pretreated with AF,AC(a pyroptosis inhibitor),or their combination for 2 h before stimulation with LPS and ATP.The changes in cell proliferation and viability were detected using CCK-8 assay,and IL-1β,IL-6,IL-18,and TNF-α levels were determined with ELISA.Immunohistochemistry,immunofluorescence assay,and immunoblotting were used to detect the protein levels of NLRP3,ASC,cleaved caspase-1,and GSDMD N in rat lung tissues and the treated cells.Results In mice with LPS exposure,AF treatment significantly improved lung pathologies and edema,reduced protein levels in BALF and pulmonary MPO level,inhibited the high expression of NLRP3/ASC/Aspase-1 axis,reduced the expression of GSDMD N,and lowered the release of IL-1β,IL-6,IL-18,and TNF-α.In RAW264.7 cells with LPS and ATP stimulation,AF pretreatment effectively reduced cell death,inhibited activation of the NLRP3/ASC/Aspase-1 axis,and reduced GSDMD N expression and the inflammatory factors.The pyroptosis inhibitor showed a similar effect to AF,and their combination produced more pronounced effects in RAW264.7 cells.Conclusion Amentoflavone can alleviate ALI in mice possibly by inhibiting NLRP3/ASC/Caspase-1 axis-mediated cell pyroptosis.关键词
穗花杉双黄酮/急性肺损伤/细胞焦亡/NLRP3/ASC/Caspase-1轴Key words
amentoflavone/acute lung injury/pyroptosis/NLRP3/ASC/Caspase-1 axis引用本文复制引用
孙亚磊,罗萌,郭长胜,高静,苏凯奇,陈立典,冯晓东..穗花杉双黄酮通过抑制细胞焦亡减轻小鼠急性肺损伤[J].南方医科大学学报,2025,45(4):692-701,10.基金项目
河南省重点研发与推广专项(科技攻关)项目(242102310508,252102310456) （科技攻关）
2023年度河南省"双一流"创建学科中医学科学研究专项(HSRP-DFCTCM-2023) （HSRP-DFCTCM-2023）
河南中医药大学博士科研启动项目(00104311-2024-2-33) （00104311-2024-2-33）
