中医学报2025，Vol.40Issue(8)：1587-1596,10.DOI:10.16368/j.issn.1674-8999.2025.08.256
金水缓纤方抑制人肺上皮细胞BEAS-2B铁死亡的活性成分及作用机制
Active Ingredient and Action Mechanism of Jinshui Huanxian Formula in Inhibiting Ferroptosis of Human Lung Epithelial Cell BEAS-2B
摘要
Abstract
Objective:To observe the effect of Jinshui Huanxian Formula(JHF)on ferroptosis of human lung epithelial cells BEAS-2B,to screen its active components,and to explore the related mechanism of action.Methods:DPPH and ABTS radical scavenging assays were used to observe the free radical scavenging activity of JHF,and calculated half maximal inhibitory concentration,IC50.Erastin was used to induce ferroptosis in BEAS-2B cells,and it was divided into blank group and model group(5 μmol·L-1 Erastin),haplomeric group(5 μmol·L-1 Erastin 10 μmol·L-1 monomer)and JHF group(5 μmol·L-1 Erastin 20 mg·L-1 JHF).CCK-8 method was used to determine cell viability,H2DCFDA reactive oxygen species(ROS)content was detected,C11-BODIPY was used to detect lip-id peroxidase(LPO)content,FerroOrange was used to detect Fe2+content,and Western blot was used to detect cystine glutamate retro-transporter(system xc-cystine/glutamate antiporter(xCT),glutathione peroxidase 4(GPX4),Ferritin protein expression levels.The active ingredient of JHF inhibiting ferroptosis was screened by offline DPPH-UHPLC-Q-Orbitrap-MS/MS technology.Results:The IC50 of Jinshui Slow Fiber Formula for DPPH and ABTS were(822.48±95.02)mg·L-1,(434.21±23.02)mg·L-1.Compared with the blank group,the cell viability of the model group was reduced(P＜0.000 1).Compared with the model group,the cell survival rate of the JHF group was increased(P＜0.000 1).Compared with the blank group,the intracellular contents of ROS,LPO and Fe2+in the model group increased(P＜0.000 1).Compared with the model group,the contents of ROS,LPO and Fe2+in the JHF group decreased(P＜0.000 1).Compared with the blank group,the protein expression levels of Ferritin,GPX4 and xCT in the model group were de-creased(P＜0.001).Compared with the model group,the protein expression levels of Ferritin,GPX4 and xCT in the JHF group in-creased(P＜0.01).The active ingredients of JHF inhibition of ferroptosis include icariin Ⅰ.,quercetin,luteolin and others.Compared with the model group,the cell survival rate of the mullein isoflavone,licochalcone A,neochlorogenic acid,hyperoside,holy herbin,quer-cetin,luteolin and icariin Ⅰ groups increased,and the contents of ROS,LPO and Fe2+decreased(P＜0.001).Compared with the model group,the protein expression levels of xCT,GPX4 and Ferritin in icariin Ⅰ group were increased(P＜0.001).Conclusion:JHF and its active ingredient icariin Ⅰ can inhibit ferroptosis in BEAS-2B cells,and its mechanism may be related to the regulation of xCT/GPX4 pathway and Ferritin pathway.关键词
肺纤维化/金水缓纤方/淫羊藿次苷Ⅰ/铁死亡/自由基/Erastin/BEAS-2B细胞/xCT/GPX4通路/Ferritin通路Key words
pulmonary fibrosis/Jinshui Huanxian Formula/Icariin Ⅰ/ferroptosis/free radical/Erastin/BEAS-2B cell/xCT/GPX4 pathway/Ferritin pathway分类
医药卫生引用本文复制引用
高艺桐,尹刘月,赵奕,钱程,张沐,赵鹏,田燕歌,刘新光..金水缓纤方抑制人肺上皮细胞BEAS-2B铁死亡的活性成分及作用机制[J].中医学报,2025,40(8):1587-1596,10.基金项目
2024年度国家科技创新2030重大专项项目(2024ZD0522905) （2024ZD0522905）
国家自然科学基金区域创新发展联合基金项目(U23A20503) （U23A20503）
河南省科技研发(优势学科)重点联合项目(232301420020) （优势学科）
