沙蟾毒精通过Nrf2-HO-1/SLC7A11途径诱导人胃癌SGC-7901细胞铁死亡  

Arenobufagin induces the ferroptosis in human gastric cancer cell line SGC-7901 through Nrf2-HO-1/SLC7A11 pathway

作　　者：王文君 刘霞霞 陈馨 程卉[2,3] 王国凯 WANG Wen-jun;LIU Xia-xia;CHEN Xin;CHENG Hui;WANG Guo-kai(School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012;Key Laboratory of Xin’an Medicine,Ministry of Education,Hefei 230038;Research Technology Center of Anhui University of Chinese Medicine,Hefei 230038)

机构地区：[1]安徽中医药大学药学院,合肥230012 [2]新安医学教育部重点实验室,合肥230038 [3]安徽中医药大学科研技术中心,合肥230038

出　　处：《中南药学》2023年第2期285-290,共6页Central South Pharmacy

基　　金：国家自然科学基金资助项目(No.81903859,No.81673650);安徽省高校自然科学研究重点项目(No.KJ2021A0591)。

摘　　要：目的 探讨沙蟾毒精(ArBu)诱导人胃癌SGC-7901细胞发生铁死亡的机制。方法 采用CCK-8法检测不同浓度ArBu对SGC-7901细胞存活率的影响,显微镜和透射电镜分别观察细胞及线粒体形态变化。将SGC-7901细胞分为对照组,ArBu低、中、高浓度组(0.2、0.4、0.8μmol·L^(-1))以及ArBu(0.4μmol·L^(-1))联合Ferrostatin-1(Fer-1,5μmol·L^(-1))组干预24 h,用试剂盒检测还原性谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、丙二醛(MDA)、总Fe和脂质活性氧(ROS)的水平。Western blot分别检测Nrf2、HO-1、SLC7A11、FTH1、GPX4的蛋白表达情况。结果 ArBu处理细胞24 h和48 h后,细胞存活率明显下降,且呈浓度依赖性。与对照组相比,ArBu处理细胞24 h后,细胞和线粒体皱缩死亡,线粒体嵴消失,GSH和SOD水平明显下降,MDA、总Fe和ROS水平明显上升。与ArBu中浓度组处理细胞24 h相比,联合Fer-1处理后,明显逆转了GSH、SOD、MDA、总Fe和脂质ROS水平,细胞形态与对照组无明显差异,线粒体并未皱缩变小。此外,ArBu明显降低Nrf2、HO-1、SLC7A11、FTH1、GPX4的蛋白表达水平。结论 ArBu能诱导人胃癌SGC-7901细胞铁死亡,其机制可能是通过Nrf2-HO-1/SLC7A11途径。Objective To determine the mechanism of ferroptosis induced by arenobufagin(ArBu) in human gastric cancer cell line SGC-7901. Methods The effect of different concentrations of ArBu on the survival rate of SGC-7901 cells was detected by CCK-8 method. Cellular and mitochondrial morphological changes were observed with microscope and transmission electron microscope,respectively. SGC-7901 cells were divided into a control group, Ar Bu low, medium, high concentration groups(0.2, 0.4 and 0.8 μmol·L^(-1)), and an ArBu(0.4 μmol·L^(-1)) combined with Ferrostatin-1(Fer-1, 5 μmol·L^(-1)) group. The intervention lasted 24 h. The levels of glutathione(GSH), superoxide dismutase(SOD), malondialdehyde(MDA), total iron(Fe) and lipid reactive oxygen species(ROS)were measured by kits. Western blot was used to detect the protein expression of Nrf2, HO-1, SLC7A11,FTH1 and GPX4. Results The cell viability decreased significantly after 24 h and 48 h of ArButreatment cells in a concentration-dependent manner. Compared with the control group, cells and mitochondria crumpled and died after 24 h of ArBu treatment, and mitochondrial cristae disappeared with a significant decrease in GSH and SOD levels, and a significant increase in MDA, total Fe and ROS levels. The levels of GSH, SOD, MDA, total Fe and lipid ROS were significantly reversed after Fer-1 combination treatment, as compared with ArBu medium concentration treatment for 24 h. The cell morphology was not significantly different from that of the control group, and the mitochondria did not crumple and shrink. In addition, Ar Bu significantly decreased the protein expression levels of Nrf2, HO-1, SLC7A11, FTH1, and GPX4. Conclusion Ar Bu can induce ferroptosis in human gastric cancer cell line SGC-7901 and the mechanism may be related to Nrf2-HO-1/SLC7A11 pathway.

关 键 词：沙蟾毒精 胃癌 铁死亡 Nrf2-HO-1/SLC7A11 

分 类 号：R285[医药卫生—中药学]