论文部分内容阅读
目的:研究亚低温对小鼠神经母细胞瘤（mouse neuro-blastoma N2a, N2a）细胞氧糖剥夺/复氧（oxygen glucose deprivation/reoxygenation, OGD/R）损伤时小泛素类修饰蛋白特异性蛋白酶3（small ubiquitin-like modifier proteins specific protease 3, SENP3）表达的影响。方法:体外培养小鼠N2a细胞并予以OGD/R处理，建立模拟大脑缺血/再灌注的OGD/R模型以及亚低温模型；将N2a细胞按照随机数字表法分为3组（每组5孔）：假手术组（S组）、OGD/R组和氧糖剥夺/复氧+亚低温组（OGD/R+HT组）。细胞计数试剂盒（cell counting kit-8, CCK-8）法检测OGD/R后N2a细胞活性，乳酸脱氢酶（lactate dehydrogenase, LDH）释放率检测OGD/R后N2a细胞毒性，Hoechst33258染色观察N2a细胞凋亡，实时荧光定量PCR（real-time quantitative PCR，RT-qPCR）分析SENP3 mRNA的相对表达量，Western blot法测定SENP3的蛋白水平。结果:与S组比较，OGD/R组和OGD/R+HT组细胞存活率下降、LDH释放率升高、凋亡细胞数量增多、SENP3 mRNA的相对表达量和蛋白水平升高（n P<0.05）；与ODG/R组比较，OGD/R+HT组细胞存活率升高、LDH释放率降低、凋亡细胞数量减少、SENP3 mRNA的相对表达量和蛋白水平降低（n P<0.05）。n 结论:亚低温减轻N2a细胞OGD/R损伤机制与抑制SENP3的表达有关。“,”Objective:To investigate the effects of mild hypothermia on the expression of small ubiquitin-like modifier proteins specific protease 3 (SENP3) in mouse neuro-blastoma (N2a) cells during oxygen-glucose deprivation/reoxygenation (OGD/R) injury.Methods:Mouse N2a cells were cultured in vitro and exposed to OGD/R to establish a model of OGD/R simulating cerebral ischemia/reperfusion and a mild hypothermia model. N2a cells were divided into three groups according to the random number table method (n n=5): group sham operation (S), group OGD/R and group OGD/R + mild hypothermia (OGD/R+HT). Cell counting kit -8 (CCK-8) assay was used to detect N2a cell viability after OGD/R, while the release rate of lactate dehydrogenase (LDH) was detected to evaluate N2a cytotoxicity after OGD/R. Hoechst 33258 staining was used to observe N2a cell apoptosis. Real-time quantitative PCR (RT-qPCR) was used to analyze the relative amounts of SENP3 mRNA. Western blot was used to measure the levels of SENP3 protein.n Results:Compared with group S, groups OGD/R and OGD/R+HT presented decreases in cell survival rate as well as increases in LDH release rate, the number of apoptotic cells, and the relative amounts of SENP3 mRNA and protein (n P<0.05). Compared with group ODG/R, group OGD/R+HT showed increases in cell survival rate and decreases in LDH release rate, the number of apoptotic neurons, and the relative amounts of SENP3 mRNA and protein (n P<0.05).n Conclusions:Mild hypothermia alleviates OGD/R injury in N2a cells, which is related with the inhibition of SENP3 expression.