时珍国医国药2026，Vol.37Issue(9)：1645-1656,12.DOI:10.70976/j.1008-0805.SZGYGY-2026-0907
补阳还五汤调控TLR4-NFκB-NLRP3炎症小体信号通路及线粒体自噬缓解LPS诱导巨噬细胞炎症的机制
Mechanism of Buyang Huanwu Decoction(补阳还五汤)on LPS-induced macrophage inflam-mation via modulating the TLR4-NF-κB-NLRP3 inflammasome pathway and mitophagy
摘要
Abstract
Objective To investigate the mechanism of Buyang Huanwu Decoction(补阳还五汤,BYHWD)in alleviating lipopoly-saccharide(LPS)-induced macrophage inflammation by modulating the TLR4-NF-κB-NLRP3 inflammasome signaling pathway and mitophagy.Methods RAW264.7 macrophages were divided into the following groups:normal,model,model control,low/medium/high-dose BYHWD,NLRP3 inhibitor(MCC950),mitophagy inhibitor(cyclosporin A),mitophagy inducer(valinomycin),and reactive oxygen species(ROS)inhibitor(N-acetylcysteine,NAC).Cell Counting Kit-8(CCK-8)assay was performed to determine cell viability and to screen optimal concentrations for LPS modeling and pharmacological interventions.Flow cytometry(FCM)was used to assess oxidative stress and ROS levels.Mitochondrial membrane potential was evaluated by JC-1 fluorescence staining.Co-localization of LysoTracker and MitoTracker probes was analyzed to examine lysosome-mitochondria fusion.The mCherry-GFP-LC3 reporter system was employed to monitor autophagosome formation and autophagic flux.NLRP3 localization and activation were detected by immunofluorescence(IF).Protein expression levels of LC3B,NLRP3,ASC,pro-caspase-1,components of the TLR4-MyD88-IRAK4-TRAF6-NF-κB pathway(including phospho-/total NF-κB and IκB),and key mitophagy regulators(NIX,BNIP3,FUNDC1,phospho-/total PINK1 and Parkin)were measured by Western blot(WB).mRNA expression of NIX,BNIP3,FUNDC1,PINK1,and Parkin was quantified by quantitative polymerase chain reaction(qPCR).Results Treatment with BYHWD-containing serum,particularly in combination with specific inhibitors,significantly reduced ROS levels(P<0.05,P<0.01)and increased the JC-1 aggregate/monomer fluorescence ratio(red/green),the ratio of red-to-yellow puncta,and the Pearson's correlation coefficient(P<0.05,P<0.01).It also upregulated the mRNA and protein expression of mitophagy-related markers(NIX,BNIP3,FUNDC1,PINK1,Parkin,LC3B,Beclin1,P62)and the ratios of phosphorylated to total PINK1 and Parkin(P<0.05,P<0.01).Conversely,the expression of TLR4,MyD88,IRAK4,TRAF6,phospho-/total NF-κB,phospho-/total IκB,NLRP3,pro-caspase-1,and ASC was downregulated(P<0.05,P<0.01).Conclusion BYHWD may alleviate LPS-induced macrophage inflammation by attenuating oxidative stress and improving mitochondrial function via suppression of the TLR4-MyD88-IRAK4-TRAF6-NF-κB pathway and modulation of mitophagy.关键词
补阳还五汤/氧化应激/巨噬细胞/动脉粥样硬化Key words
Buyang Huanwu Decoction(补阳还五汤)/Oxidative stress/Macrophages/Atherosclerosis分类
医药卫生引用本文复制引用
贺智贤,常诗瑶,俞虹,游宇,刘玉晖..补阳还五汤调控TLR4-NFκB-NLRP3炎症小体信号通路及线粒体自噬缓解LPS诱导巨噬细胞炎症的机制[J].时珍国医国药,2026,37(9):1645-1656,12.基金项目
国家自然科学基金(82360785),江西省自然科学基金重点项目(20232ACB206058),江西省研究生创新专项资金项目(YC2025-S208) （82360785）
