摘要
目的探讨白藜芦醇(Res)对1-甲基-4-苯基吡啶(MPP^(+))诱导的SH-SY5Y细胞内铁死亡及氧化应激的影响。方法选择MPP^(+)处理SH-SY5Y细胞来建立帕金森病(PD)的细胞模型,共分为4组:CTL组为不做处理组;Res组为使用浓度为25μmol/L的Res处理细胞;MPP^(+)组为使用浓度为500μmol/L的MPP^(+)处理细胞;Res+MPP^(+)组为先用浓度为25μmol/L的Res处理细胞,1 h后再加入MPP^(+)处理细胞。使用噻唑蓝(MTT)法检测4组细胞存活率及细胞毒性,分别测定4组细胞的活性氧(ROS)、线粒体膜电位及Fe^(2+)水平。免疫印记法测定4组细胞的谷胱甘肽过氧化物酶(GPX4)及胱氨酸/谷氨酸逆向转运蛋白(SLC7A11)蛋白表达水平。结果与CTL组相比,500μmol/L浓度的MPP^(+)细胞存活率降低约一半,差异有统计学意义(t=17.15,P<0.01)。与MPP^(+)组相比,Res+MPP^(+)组的细胞存活率显著增加,差异有统计学意义(t=3.13,P<0.01)。与MPP^(+)组相比,Res+MPP^(+)组的细胞毒性显著降低,差异有统计学意义(t=3.20,P<0.01)。与MPP^(+)组相比,Res+MPP^(+)组的ROS水平及Fe^(2+)水平显著降低,四甲基罗丹明乙酯(TMRE)荧光强度显著增加,差异均有统计学意义(t=5.26、3.80、4.72,P均<0.05)。与MPP^(+)组相比,Res+MPP^(+)组的GPX4蛋白表达水平及SLC7A11蛋白表达水平显著增加,差异均有统计学意义(t=5.10、2.84,P均<0.05)。结论Res可以通过提高GPX4及SLC7A11蛋白表达水平来抑制MPP^(+)诱导的SH-SY5Y细胞内的铁死亡,进而减轻氧化应激并发挥保护细胞的作用。
Objective To investigate the effects of resveratrol(Res)on ferroptosis and oxidative stress in SH-SY5Y cells induced by 1-methyl-4-phenylpyridinium(MPP^(+)).Methods SH-SY5Y cells were selected to establish a cell model of Parkinson's disease(PD)by MPP^(+),and the cells were divided into 4 groups:CTL group was the untreated group;Res group was treated with 25μmol/L Res;MPP group was treated with MPP^(+)at a concentration of 500μmol/L;the Res+MPP^(+)group was treated with Res at a concentration of 25μmol/L for 1 h,and then MPP^(+)was added to treat the cells.The methylthiazolyldiphenyl-tetrazolium bromide(MTT)method was used to detect the cell viability and cytotoxicity of 4 groups of cells,and the reactive oxygen species(ROS),mitochondrial membrane potential and Fe2 levels of 4 groups of cells were measured.The immunomarkers assay was used to determine the expression levels of glutathione peroxidase(CPX4)and solute carrier family 7-member 11(SLC7A11)in 4 groups of cells.ResulstsCompared with the CTLgroup,the cell survival rate of MPP^(+)at 500μmol/L was reduced by about half,and the difference was statistically significant(t=17.15,P<0.01).Compared with the MPP^(+)group,the cell survival rate of Res+MPP^(+)group was significantly increased,and the difference was statistically significant(t=3.13,P<0.01).Compared with the MPP^(+)group,the cytotoxicity of Res+MPP^(+)group was significantly reduced,and the difference was statistically significant(t=3.20,P<0.01).Compared with the MPP^(+)group,the ROS level and Fe^(2+)level in the Res+MPP group were significantly reduced,and the tetramethyl rhodamine ethyl ester(TMRE)fluorescence intensity was significantly increased(t=5.26,3.80,4.72;all P<0.05).Compared with the MPP group,the protein expression levels of CPX4 and SLC7A11 in the Res+MPP^(+) group were significantly increased(t=5.10,2.84;both P<0.05).Conclusions Res could inhibit MPP^(+)-induced ferroptosis in SH-SY5Y cells by increasing the protein expression levels of GPX4 and SLC7A11,thereby alleviating oxidative stress and protecting cells.
作者
钟佳宏
余玺辉
练昌林
钟运鸣
林卓苗
ZHONG Jiahong;YU Xihui;LIAN Changlin;ZHONG Yunming;LIN Zhuomiao(Department of Clinical Pharmacy,Meizhou People's Hospital,Meizhou,Guangdong 514031,China;Department of Pharmacy,the Second Affiliated Hospital of Shantou University Medical College,Shantou,Guangdong 515000,China;Department of Emergency,Guangdong Provincial People's Hospital,Guangzhou,Guangdong 510000,China)
出处
《热带医学杂志》
2025年第8期1017-1021,共5页
Journal of Tropical Medicine
基金
国家自然科学基金(82301436)
广东省基础与应用基础研究基金联合基金(2023A1515111199)
梅州市社会发展科技计划项目(2023B05,2023B06)。
