摘要
目的探讨鱼藤酮干预人神经母细胞瘤细胞(SH-SY5Y)后细胞内氧化物和抗氧化物的表达水平。方法常规胎牛血清培养2 d后,根据不同细胞处理方式将细胞分为正常组和鱼藤酮组,其中鱼藤酮组按照浓度梯度分亚组为25、50、100、200、500、1000、2000 nM(nM:nmol/L)。分别使用光镜观察上述不同鱼藤酮干预浓度梯度细胞形态学变化以及MTT比色法检测细胞存活和生长,从而探讨最佳鱼藤酮干预浓度。使用流式细胞仪观察鱼藤酮干预后的细胞内氧化水平DCFH-DA变化;采用酶联免疫吸附法测定细胞内超氧化物歧化酶(SOD)活性水平以及细胞内氧化物质脂质过氧化水平。结果MTT检测和光镜检测显示,SH-SY5Y细胞活力与鱼藤酮的干预浓度呈浓度依赖性,诱发细胞氧化应激损伤而不过度造成细胞死亡的浓度为500 nM的鱼藤酮,而2000 nM的鱼藤酮加入培养基后细胞几乎停止生长。流式细胞术和酶联免疫吸附法检测使用500 nM鱼藤酮处理后的DCFH-DA染色SH-SY5Y细胞内的氧化应激水平增高,抗氧化物SOD活性水平减少,氧化物脂质过氧化产物(TBARS)增多。结论鱼藤酮促进SH-SY5Y细胞内抗氧化物SOD活性水平减少,氧化物脂质过氧化产物增多,氧化应激水平增高。SH-SY5Y细胞活力与鱼藤酮的干预浓度呈浓度依赖性,浓度越高细胞死亡越多。
Objective To investigate the expression of intracellular oxides and antioxidants in human neuroblastoma cells(SH-SY5Y)effected by rotenone.Methods The cells were divided into normal group and rotenone group according different treatment after two days of conventional fetal bovine serum culture.The rotenone group was divided into five subgroups according to the concentration gradient of 25,50,100,200,500,1000,2000 nM(nM:nmol/L).The morphological changes of the cells were observed by light microscope and the survival and growth of the cells were detected by MTT colorimetry.Flow cytometry was used to observe the changes of DCFH-DA in the cells after rotenone intervention,and enzyme-linked immunosorbent assay was used to determine the activity of superoxide dismutase(SOD)and lipid peroxidation.Results It was confirmed that the cell viability of SH-SY5Y was dose-dependent with the intervention concentration of rotenone by MTT assay and light microscopy detection.The concentration of 500 nM rotenone could induce oxidative stress injury without excessive cell death,while the growth of cells almost stopped when 2000 nM rotenone was added into the medium.Flow cytometry and enzyme-linked immunosorbent assay showed that DCFH-DA staining of SH-SY5Y cells treated with 500 nM rotenone increased the level of oxidative stress,decreased the activity of SOD and increased the level of oxide lipid peroxidation products(TBARS).Conclusion Rotenone could regulate down of SOD activity in SH-SY5Y cells,increase the production of lipid peroxidation and increase the level of oxidative stress.The cell viability of SH-SY5Y was positively correlated with the concentration of rotenone.The higher the concentration,the more cell death.
作者
张萍
邵靓
ZHANG Ping;SHAO Liang(Department of Cardiology,Jiangxi Provincial People′s Hospital,Jiangxi Province,Nanchang,330000,China;Department of Neurology,Jiangxi Provincial People′s Hospital,Jiangxi Province,Nanchang,330000,China)
出处
《中国当代医药》
CAS
2021年第20期8-11,共4页
China Modern Medicine
基金
浙江省自然科学基金青年基金项目(LQ18H090004)
江西省青年科学基金项目(20202BAB216014)
江西省卫生健康委科技计划项目(202110005)。
