仙鹤草素通过抑制PI3K-AKT通路促进NRF2介导的胃癌细胞铁死亡  

Agrimoiin promotes NRF2-mediated ferroptosis of gastric cancer cells by inhibiting the PI3KAKT pathway

作　　者：杨明月 董鑫 冷孟桐 迟文成[4] 姜家康[4] YANG Mingyue;DONG Xin;LENG Mengtong;CHI Wencheng;JIANG Jiakang(Heilongjiang University of Chinese Medicine,Harbin 150000,China;Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Tianjin University of Traditional Chinese Medicine,Tianjin 301600,China;Department of Oncology,the First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150000,China)

机构地区：[1]黑龙江中医药大学,哈尔滨150000 [2]广州中医药大学,广州510405 [3]天津中医药大学,天津301600 [4]黑龙江中医药大学附属第一医院肿瘤科,哈尔滨150000

出　　处：《广西医科大学学报》2024年第7期1017-1024,共8页Journal of Guangxi Medical University

基　　金：国家自然科学基金面上项目(No.82305114);黑龙江中医药大学“双一流”中西医结合学科发展助力基金(No.HLJSYL21002)。

摘　　要：目的:观察仙鹤草素抑制人胃癌HGC-27、MKN-45的作用机制。方法:将胃癌HGC-27、MKN-45细胞分为对照组、不同剂量仙鹤草素组、卡培他滨(阳性对照)组及仙鹤草素+NRF2激活剂(Bardoxolone)组。采用CCK-8法检测HGC-27、MKN-45细胞活力,划痕实验观察细胞迁移能力,流式细胞术检测细胞内活性氧(ROS)水平,酶联免疫吸附试验(ELISA)法检测细胞丙二醛(MDA)、谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)水平,western blotting法检测PI3K-AKT通路相关蛋白及抗铁死亡分子NRF2、Histone H3、HO-1、SLC7A11、GPX4表达。结果:与对照组相比,不同剂量仙鹤草素对HGC-27、MKN-45细胞活性均有显著的抑制作用(P<0.05),且随着时间延长和仙鹤草素剂量增加,抑制作用增强。与对照组比较,仙鹤草素低、高剂量组和阳性对照组HGC-27、MKN-45细胞迁移能力减弱,ROS和MDA水平升高,GSH/GSSG、P-PI3K/PI3K、P-AKT/AKT比值降低,总NRF2(Total-NRF2)、细胞核内NRF2(Nuclear-NRF2)、HO-1、SLC7A11、GPX4蛋白表达下调(均P<0.05)。仙鹤草素+Bardoxolone组Total-NRF2、GPX4蛋白表达水平升高(P<0.05)。结论:仙鹤草素可促进胃癌HGC-27、MKN-45细胞铁死亡,其作用机制可能与抑制PI3K-AKT-NRF2信号通路的激活有关。Objective:To observe the inhibitory mechanism of agrimoiin on human gastric cancer cell line HGC-27 and MKN-45.Methods:Gastric cancer HGC-27 and MKN-45 cells were divided into control group,different doses of agrimoniin groups,capecitabine(positive control)group and agrimoniin+NRF2 activator(Bardoxolone)group.Cell counting kit-8(CCK-8)assay was used to detect the viability of HGC-27 and MKN-45 cells.Scratch test was used to observe the cell migration ability.Flow cytometry was used to detect the level of intracellular re-active oxygen species(ROS).Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of malondialdehyde(MDA),glutathione(GSH)and oxidized glutathione(GSSG).Western blotting was used to de-tect the expression of PI3K-AKT pathway-related proteins and anti-ferroptosis molecules NRF2,Histone H3,HO-1,SLC7A11 and GPX4.Results:Compared with the control group,different doses of agrimoniin had a signifi-cant inhibitory effect on the activity of HGC-27 and MKN-45 cells(P<0.05),and the inhibitory effect increased with the extension of time and the increase of agrimoniin dose.Compared with the control group,the migration ability of HGC-27 and MKN-45 cells in the low-dose and high-dose agrimoniin groups and the positive control group was weakened,the levels of ROS and MDA were increased,the ratios of GSH/GSSG,P-PI3K/PI3K,and P-AKT/AKT were decreased,and the expression of Total-NRF2,nuclear-NRF2,HO-1,SLC7A11,and GPX4 pro-teins was down-regulated(all P<0.05).The protein expression levels of Total-NRF2 and GPX4 were increased in the agrimoniin+Bardoxolone group(P<0.05).Conclusion:Agrimoniin can promote ferroptosis in gastric can-cer HGC-27 and MKN-45 cells,and its mechanism may be related to inhibiting the activation of PI3K-AKT-NRF2 signaling pathway.

关 键 词：仙鹤草素 PI3K-AKT信号通路 胃癌 铁死亡 

分 类 号：R735.2[医药卫生—肿瘤]