曲克芦丁通过TP53抑制Erastin诱导的人正常结肠上皮细胞和结直肠癌细胞铁死亡  

Troxerutin inhibits Erastin-induced ferroptosis in human normal colonic epithelial cells and colorectal cancer cells via TP 53

作　　者：刘付翠娟 蒋代顺 张璐 陈帅 叶华 夏海珊 刘义 LIU-FU Cuijuan;JIANG Daishun;ZHANG Lu;CHEN Shuai;YE Hua;XIA Haishan;LIU Yi(School of Pharmacy,Guangdong Medical University,Dongguan Guangdong 523808;First Dongguan Affiliated Hospital,Guangdong Medical University,Dongguan Guangdong 523710;School of Ocean and Tropical Medicine,Guangdong Medical University,Zhanjiang Guangdong 524023,China)

机构地区：[1]广东医科大学药学院,广东东莞523808 [2]广东医科大学附属东莞第一医院,广东东莞523710 [3]广东医科大学海洋与热带医学学院,广东湛江524023

出　　处：《临床与病理杂志》2024年第9期1177-1190,共14页Journal of Clinical and Pathological Research

基　　金：广东省自然科学基金(2018A0303130252);湛江市科技发展专项项目(2019A01019)。

摘　　要：目的:铁死亡被证实在结直肠癌发生、发展和治疗中发挥重要作用。曲克芦丁具有多种生物学活性,但其是否与铁死亡有关还不得而知。本研究旨在探讨曲克芦丁对人正常结肠上皮细胞和结直肠癌细胞铁死亡的影响。方法:采用噻唑蓝(methylthiazolyldiphenyl-tetrazolium bromide,MTT)法检测曲克芦丁对铁死亡诱导剂Erastin诱导的结肠上皮细胞及结直肠癌细胞的细胞活力的影响,流式细胞仪检测曲克芦丁对细胞内脂质过氧化物、亚铁离子水平的影响,比色法检测细胞内丙二醛(malondialdehyde,MDA)及还原型谷胱甘肽(reduced glutathione,GSH)/氧化型谷胱甘肽(oxidized glutathione disulfide,GSSG)比值的变化,荧光染色法检测细胞线粒体膜电位。使用网络药理学和分子对接技术预测曲克芦丁参与铁死亡的潜在靶点,并采用蛋白质印迹法对其潜在靶点进行验证。结果:曲克芦丁能有效逆转由Erastin诱导的结肠上皮细胞和结直肠癌细胞铁死亡,并能显著降低由Erastin引起的细胞内脂质过氧化物、亚铁离子和MDA水平升高,提高细胞内GSH/GSSG比值、线粒体膜电位及胱氨酸/谷氨酸逆向转运蛋白溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)和谷胱甘肽过氧化酶4(glutathione peroxidase 4,GPX4)的表达水平;网络药理学、分子对接及蛋白质印迹法检测结果显示肿瘤蛋白p53(tumor protein p53,TP53)可能是曲克芦丁抑制结肠上皮细胞及结直肠癌细胞铁死亡的潜在靶点。结论:曲克芦丁通过降低TP53水平来抑制结肠上皮细胞和结直肠癌细胞的铁死亡。Objective:Ferroptosis has been shown to play a significant role in the onset,progression,and treatment of colorectal cancer.Troxerutin possesses diverse biological activities,but its relationship with ferroptosis remains unclear.This study aims to investigate the effects of troxerutin on ferroptosis in human normal colonic epithelial cells and colorectal cancer cells.Methods:The effects of troxerutin on cell viability were assessed using the methylthiazolyldiphenyl-tetrazolium bromide(MTT)assay in Erastin-induced ferroptosis in colonic epithelial cells and colorectal cancer cells.Flow cytometry was used to evaluate intracellular lipid peroxides and ferrous ion levels.Colorimetric assays were conducted to measure intracellular malondialdehyde(MDA)and the reduced glutathione(GSH)/oxidized glutathione disulfide(GSSG)ratio.Mitochondrial membrane potential was assessed using fluorescence staining.Network pharmacology and molecular docking were employed to predict potential ferroptosis-related targets of troxerutin,which were subsequently validated by Western blotting.Results:Troxerutin effectively reversed Erastin-induced ferroptosis in both colonic epithelial cells and colorectal cancer cells.It significantly reduced Erastin-induced elevations in lipid peroxides,ferrous ions,and MDA levels,while increasing the intracellular GSH/GSSG ratio,mitochondrial membrane potential,and the expression levels of solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4).Network pharmacology,molecular docking,and western blotting results suggested that tumor protein p53(TP53)might be a potential target of troxerutin in inhibiting ferroptosis.Conclution:Troxerutin inhibits ferroptosis in colonic epithelial cells and colorectal cancer cells by downregulating TP53 levels.

关 键 词：曲克芦丁 铁死亡 结肠上皮细胞 结直肠癌细胞 肿瘤蛋白p53 

分 类 号：R73[医药卫生—肿瘤]