摘要
目的 研究京尼平苷(Gen)对氧糖剥夺/复氧(OGD/R)诱导的人神经母细胞瘤细胞SH-SY5Y损伤的保护作用,并探讨其机制.方法 将SH-SY5Y细胞分为对照组、OGD/R组及Gen低、中、高浓度组(12.5、25、50 μmol·L-1).除对照组外,其他各组细胞均建立OGD/R损伤模型,并采用不同浓度Gen进行干预.CCK-8检测细胞增殖活性;生化法检测细胞上清液乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽(GSH)水平;比色法检测细胞内Fe2+水平;流式细胞术检测细胞内活性氧(ROS)水平;Western blot检测细胞内谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(SLC7A11)和转铁蛋白受体1(TFR1)蛋白表达水平.结果 与对照组比较,OGD/R组细胞LDH释放率、MDA含量、Fe2+水平、ROS水平和TFR1蛋白表达水平显著升高(P<0.05),而细胞增殖活性、SOD活性、GSH水平、SLC7A11和GPX4蛋白表达水平显著降低(P<0.05).与OGD/R组比较,Gen各浓度组细胞LDH释放率、MDA含量、Fe2+水平、ROS水平和TFR1蛋白表达水平显著降低(P<0.05),而细胞增殖活性、SOD活性、GSH水平、SLC7A11和GPX4蛋白表达水平显著升高(P<0.05),其中以H-Gen组最为显著(P<0.05).结论 Gen可抑制OGD/R诱导的SH-SY5Y细胞氧化应激与铁死亡,其作用机制可能与激活SLC7A11/GPX4信号通路有关.
Abstract
Objective To study the effects of geniposide(Gen)on the injury of human neuroblastoma cell SH-SY5Y induced by oxygen glucose deprivation/reoxygenation(OGD/R)and its mechanism.Methods SH-SY5Y cells were divided into control group,OGD/R group,low medium and high concentration groups of Gen(12.5,25,50 μmol·L-1).Except the control group,OGD/R injury models were established in other groups,and different concentrations of Gen were used for intervention.Cell proliferation activity was detected by CCK-8 assay.The levels of lactate dehydrogenase(LDH),superoxide dismutase(SOD),malonaldehyde(MDA)and glutathione(GSH)in cell supernatant were detected by biochemical method.The intracellular Fe2+ level was detected by colorimetry.The level of reactive oxygen species(ROS)in cells was detected by flow cytometry.The protein expression levels of glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11)and transferrin receptor 1(TFR1)in cells were detected by Western blot.Results Compared with the control group,LDH release rate,MDA content,Fe2+ level,ROS level and TFR1 expression level in OGD/R group were significantly increased(P<0.05),while cell proliferation activity,SOD activity,GSH level,SLC7A11 and GPX4 expression levels were significantly decreased(P<0.05).Compared with the OGD/R group,the LDH release rate,MDA content,Fe2+ level,ROS level and TFR1 expression level of cells in each concentration group of Gen decreased significantly(P<0.05),while cell proliferation activity,SOD activity,GSH level,SLC7A11 and GPX4 expression levels increased significantly(P<0.05),especially in H-Gen group.Conclusion Gen can inhibit oxidative stress and ferroptosis of SH-SY5Y cells induced by OGD/R,and its mechanism may be related to the activation of SLC7A11/GPX4 signal pathway.
基金项目
贵州省科技厅(贵州省知识产权局)基础研究计划(黔科合基础[2017]1213)
遵义市科技局科技合作项目(遵市科合HZ字2021240号)
国家科技期刊平台
NSTL
万方数据