消瘤藤总香豆素通过诱导铁死亡抑制小细胞肺癌H1688细胞增殖  

Total coumarins in Pileostegia tomentella inhibits proliferation of small cell lung cancer H1688 cells by inducing ferroptosis

作　　者：黎骊 陆国寿 王丽 吕纪华 黄建猷 刘瑛 LI Li;LU Guo-shou;WANG Li;LYU Ji-hua;HUANG Jian-you;LIU Ying(Guangxi Key Laboratory of Traditional Chinese Medicine Quality Standards,Guangxi Institute of Chinese Medical and Pharmaceutical Sciences,Nanning 530022,China)

机构地区：[1]广西中药质量标准研究重点实验室,广西壮族自治区中医药研究院,广西南宁530022

出　　处：《中国药理学通报》2023年第6期1115-1121,共7页Chinese Pharmacological Bulletin

基　　金：广西自然科学基金青年科学基金项目(No 2022GXNS FBA035500);广西自然科学基金面上项目(No 2020GX NSFAA259067);广西壮族自治区中医药研究院自主研究项目(No桂中自研2021003)。

摘　　要：目的研究消瘤藤总香豆素(total coumarins from Pileostegia tomentella,TCPT)通过诱导铁死亡抑制小细胞肺癌(small cell lung cancer,SCLC)H1688细胞增殖的机制。方法采用梯度浓度的TCPT处理SCLC细胞H1688后,CCK-8法检测TCPT对细胞增殖的抑制作用;利用透射电镜观察给药后线粒体形态学变化;二氯荧光素(DCFH-DA)荧光探针检测细胞内活性氧(ROS)水平;BODIPY 581/589 C11探针检测细胞内脂质过氧化物的生成能力;Western blot检测细胞中铁死亡相关蛋白谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)、Kelch样环氧氯丙胺相关蛋白1(Kelch-like ECH-associated protein,KEAP1)、核因子E2相关因子2(nuclear factor E2 related factor 2,NRF2)、铁蛋白重链1(ferritin heavy chain 1,FTH1)的表达水平。结果TCPT干预SCLC细胞H1688后,对细胞增殖有明显抑制作用(P<0.05,P<0.01)。透射电镜下可见TCPT引起SCLC铁死亡形态学变化。TCPT可有效提高细胞内ROS和脂质过氧化物水平。H1688中铁死亡标志蛋白GPX4、NRF2、FTH1的表达下调而KEAP1的表达上调,差异有统计学意义(P<0.01)。结论PCPT可明显抑制SCLC细胞H1688增殖能力,其机制可能是通过升高细胞内ROS和脂质过氧化水平,最终诱导细胞铁死亡。Aim To explore the mechanism by which total coumarins in Pileostegia tomentella(TCPT)inhibits the proliferation of small cell lung cancer(SCLC)H1688 cells via inducing ferroptosis.Methods The gradient concentrations of TCPT were used to treat H1688 cells.CCK-8 assay was applied for detection of proliferative inhibition of H1688 cells.Trans-mission electron microscopy was used to approach the morphological changes of H1688 cells under the treatment of TCPT.Additionally,dichlorofluorescein(DCFH-DA)probe was used to detect the intracellular reactive oxygen species(ROS)level.BODIPY 581/589 C11 probe was applied to examine the intracellular lipid peroxide formation.Western blotting was employed to detect the expression levels of glutathione peroxidase 4(GPX4),kelch-like ECH-associated protein(KEAP1),nuclear factor E2 related factor 2(NRF2),ferritin heavy chain 1(FTH1)proteins in H1688 cells.Results The proliferation of small cell lung cancer cell H1688 was dramatically inhibited after TCPT intervention(P<0.05,P<0.01).The morphological characteristics of ferroptosis induced by TCPT were observed by transmission electron microscope.TCPT could also effectively elevate intracellular level of ROS and lipid peroxide.In H1688 cells the expression of ferroptosis markers GPX4,NRF2,and FTH1 was down-regulated,while the expression of KEAP1 was up-regulated,and there were statistically significant differences among the markers mentioned above(P<0.01).Conclusions Total coumarins in TCPT can significantly inhibit the proliferation of H1688 cells,possibly through increasing ROS and intracellular lipid peroxide levels and eventually inducing ferroptosis.

关 键 词：小细胞肺癌 消瘤藤 总香豆素 脂质过氧化 铁死亡 细胞增殖 

分 类 号：R284.1[医药卫生—中药学] R329.28[医药卫生—中医学] R591.1R734.2