摘要
目的:研究甘草水提物能否促进Erastin诱导结直肠癌(CRC)细胞株发生铁死亡。方法:2种CRC细胞(Caco-2、SW837)分别分为5组:空白对照组、甘草水提物低浓度(50μg/mL)组、甘草水提物高浓度(100μg/mL)组、甘草水提物高浓度(100μg/mL)+铁死亡诱导剂Erastin(30μmol/L)组、甘草水提物高浓度(100μg/mL)+铁死亡诱导剂Erastin(30μmol/L)+Fer-1(30 nmol/L)组。MTT检测细胞活力,流式细胞术检测细胞凋亡、死亡情况及细胞ROS生成量、线粒体膜电位变化,CCK-8实验验证细胞死亡方式,并检测Fe^(2+)、MDA、GSH水平和SLC7A11、DPP4、NOX1、GPX4蛋白表达。结果:与空白对照组比较,程序性坏死性凋亡抑制剂Necrostatin-1、凋亡抑制剂Z-VAD-FMK、自噬抑制剂3-MA均不能阻断甘草水提物联合Erastin处理导致的细胞死亡,而铁死亡抑制剂Fer-1能抑制甘草水提物联合Erastin处理导致的细胞死亡,说明细胞死亡为铁死亡。甘草水提物100μg/mL组和甘草水提物100μg/mL+Erastin组Caco-2、SW837细胞中ROS生成量、MDA含量、Fe^(2+)含量及DPP4蛋白表达显著升高,线粒体膜电位、GSH水平及SLC7A11、NOX1、GPX4蛋白表达显著降低(P<0.05或P<0.01)。与甘草水提物100μg/mL+Erastin组比较,Fer-1的加入使得Caco-2、SW837细胞中ROS生成量、MDA含量、Fe^(2+)含量及DPP4蛋白表达显著降低,线粒体膜电位、GSH水平及SLC7A11、NOX1、GPX4蛋白表达显著升高(P<0.05或P<0.01)。结论:甘草水提物能诱导CRC细胞株发生铁死亡。
Objective:To investigate whether Glycyrrhizae Radix et Rhizoma water extracts could promote iron death in Erastin treated colorectal cancer(CRC)cells.Methods:Two kinds of CRC cells(Caco-2 and SW837)were randomly divided into 5 groups,including normal control group,Glycyrrhizae Radix et Rhizoma water extract low(50μg/mL),high(100μg/mL)concentration groups,Glycyrrhizae Radix et Rhizoma water extract high concentration(100μg/mL)with iron death attractor Erastin(30μmol/L)group,Glycyrrhizae Radix et Rhizoma water extract high concentration(100μg/mL)with Erastin(30μmol/L)and Fer-1(30 nmol/L)group.Cell viability was determined by MTT,the cell apoptosis,death,and ROS production,mitochondrial membrane potential changes were determined by flow cytometry,cell death was verified by CCK-8 assay,Fe^(2+),MDA,GSH levels and SLC7A11,DPP4,NOX1,GPX4 protein expressions were measured.Results:Compared with normal control group,programmed necrotizing inhibitor of apoptosis Necrostatin-1,apoptosis inhibitor Z-VAD-FMK and autophagy inhibitor 3-MA could not block cell death induced by Glycyrrhizae Radix et Rhizoma water extract combined with Erastin treatment.Ferroptosis inhibitor Fer-1 inhibited cell death caused by Glycyrrhizae Radix et Rhizoma water extract combined with Erastin,indicating that cell death was iron death.ROS production,MDA content,Fe^(2+)content and DPP4 protein expression in Caco-2 and SW837 cells of Glycyrrhizae Radix et Rhizoma water extract(100μg/mL)group and Glycyrrhizae Radix et Rhizoma water extract(100μg/mL)with Erastin group were significantly increased.Mitochondrial membrane potential,GSH level and SLC7A11,NOX1,GPX4 protein expressions were significantly decreased(P<0.05 or P<0.01).Compared with Glycyrrhizae Radix et Rhizoma water extract(100μg/mL)with Erastin group,the addition of Fer-1 significantly decreased ROS production,MDA content,Fe^(2+)content and DPP4 protein expression in Caco-2 and SW837 cells.Mitochondrial membrane potential,GSH level and SLC7A11,NOX1,GPX4 protein expressions were significantly increased(P<0.05 or P<0.01).Conclusion:Glycyrrhizae Radix et Rhizoma water extract can induced iron death in CRC cell lines.
作者
尚韬
王城磊
庞石凯
SHANG Tao;WANG Cheng-lei;PANG Shi-kai(The First Affiliated Hospital of Zhejiang Chinese Medicine University/Zhejiang Hospital of Traditional Chinese Medicine,Hangzhou 310020,China)
出处
《中药材》
CAS
北大核心
2023年第5期1261-1266,共6页
Journal of Chinese Medicinal Materials
基金
浙江省中医药科技计划项目(2021ZB095)
关键词
甘草水提物
Erastin
结直肠癌
铁死亡
Glycyrrhizae Radix et Rhizoma water extracts
Erastin
Colorectal cancer(CRC)
Iron death
作者简介
尚韬(1988-),男,硕士,主治医师,研究方向:肛肠外科,Tel:13588086366,E-mail:shangtaomail@163.com;通讯作者:庞石凯,E-mail:psk72@126.com。
