锌指蛋白580过表达对氧糖剥夺SH-SY5Y细胞凋亡的影响及可能机制  

Effect of overexpressed zinc finger protein 580 on apoptosis of oxygen-glucose deprivation SH-SY5Y cells and its possible mechanism

作　　者：阴崇娟[1] 阴怀清[1] 栗红[1] 白丹 夏莉[1] 郭晋伟[1] 刘文俊 张新[1] Yin Chongjuan;Yin Huaiqing;Li Hong;Bai Dan;Xia Li;Guo Jinwei;Liu Wenjun;Zhang Xin(Department of Pediatrics,First Hospital of Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西医科大学第一医院儿科,太原030001

出　　处：《中华围产医学杂志》2022年第11期838-843,共6页Chinese Journal of Perinatal Medicine

基　　金：山西省回国留学人员科研项目(2017-113);山西省应用基础研究计划项目(20210302124650);山西省卫生健康委科研计划项目(2022027)。

摘　　要：目的探讨锌指蛋白580(zinc finger protein 580,ZNF580)在SH-SY5Y细胞系氧糖剥夺(oxygen-glucose deprivation,OGD)模型中的表达情况及其过表达对缺氧缺血神经元凋亡的影响及可能机制。方法研究分两部分:(1)培养人神经母细胞瘤SH-SY5Y细胞系,分为模型组和对照组,模型组建立OGD模型(95%N_(2)、5%CO_(2)、0.1%O2的三气培养箱37℃恒温培养6 h),并于OGD后6、12和24 h提取蛋白,利用蛋白质印迹技术定量检测ZNF580的表达情况。(2)慢病毒转染过表达ZNF580对细胞凋亡及半胱氨酸蛋白酶3的活化形式(cleaved caspase-3)表达的影响:取对照组和模型组OGD后24 h的细胞。过表达组细胞首先通过慢病毒转染技术构建过表达ZNF580细胞,再行OGD处理。采用流式细胞技术检测各组细胞凋亡率,采用蛋白质印迹技术检测cleaved caspase-3的表达。采用两独立样本t检验、单因素方差分析及LSD-t两两比较进行统计学分析。结果(1)与对照组(1.00)相比,模型组OGD后6、12及24 h ZNF580相对表达量分别为1.36±0.05、2.12±0.07和1.69±0.05,均显著升高(LSD-t值分别为9.20、28.26和19.21,P值均<0.001)。(2)对照组、模型组、过表达组细胞凋亡率分别为(1.07±0.56)%、(21.51±1.65)%和(3.42±0.93)%,cleaved caspase-3相对表达量分别为1.00、2.47±0.59和1.70±0.25。与对照组相比,模型组的细胞凋亡率及cleaved caspase-3相对表达量升高(LSD-t值分别为21.98和8.17,P值均为0.001);与模型组相比,过表达组的细胞凋亡率及cleaved caspase-3相对表达量降低(LSD-t=19.45,P=0.001;LSD-t=4.28,P=0.005)。结论缺氧缺血可导致ZNF580过表达,ZNF580过表达可能通过抑制cleaved caspase-3表达从而影响其酶解活化来减轻缺氧缺血神经元的凋亡。Objective To investigate the expression of zinc finger protein 580(ZNF580)in oxygen-glucose deprivation(OGD)model of SH-SY5Y cell line and its overexpression on the apoptosis of hypoxic-ischemic neurons and the possible mechanism.Methods The study was divided into two parts:(1)Human neuroblastoma SH-SY5Y cell line was cultured and divided into the model group and control group.The model group was incubated at 37℃for 6 h in a three-gas incubator of 95%N2,5%CO_(2),and 0.1%O_(2) to establish OGD model,and proteins were extracted at 6,12,and 24 h after OGD.The expression of ZNF580 was quantified by Western blot.(2)Effects of ZNF580 overexpressed with lentivirus transfection on the apoptosis and cleaved caspase-3 expression:Cells were collected from the control group and model group 24 h after OGD.Overexpressed ZNF580 cells were constructed by lentivirus transfection as the overexpression group and then treated with OGD.Flow cytometry was used to detect the apoptosis rate in the three groups and Western blot was used to detect the expression of cleaved caspase-3.Two independent sample t-test,one-way variance analysis,and LSD-t for pairwise comparison were used for statistical analysis.Results(1)ZNF580 expression was significantly increased at 6,12,and 24 h after OGD compared with the control group(1.36±0.05,2.12±0.07,1.69±0.05 vs 1.00,LSD-t=9.20,28.26,and 19.21,all P<0.001).(2)Apoptosis rates of the control,model,and overexpression groups were(1.07±0.56)%,(21.51±1.65)%,and(3.42±0.93)%,respectively,and relative expression levels of cleaved caspase-3 were 1.00,2.47±0.59,and 1.70±0.25,respectively.Compared with the control group,apoptosis rate and cleaved caspase-3 relative expression level were significantly increased in the model group(LSD-t=21.98 and 8.17,both P=0.001),while the two figures were significantly decreased in the overexpression group when compared with the model group(LSD-t=19.45,P=0.001;LSD-t=4.28,P=0.005).Conclusion Hypoxia and ischemia could lead to the overexpression of ZNF580,which may redu

关 键 词：转录因子 神经元 细胞凋亡 半胱氨酸天冬氨酸蛋白酶3 缺氧缺血 脑 细胞系 肿瘤 慢病毒属 转染 

分 类 号：R743[医药卫生—神经病学与精神病学]