摘要
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(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(P<0.05)。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=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.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(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(P<0.05).Conclusions Mild hypothermia alleviates OGD/R injury in N2a cells,which is related with the inhibition of SENP3 expression.
作者
孙贵亮
徐玉云
王培
张高峰
王明山
Sun Guiliang;Xu Yuyun;Wang Pei;Zhang Gaofeng;Wang Mingshan(Graduate School of Qingdao University,Qingdao 266071,China;Department of Cadre Health Care,Qingdao Municipal Hospital,Qingdao 266071,China;Graduate School of Dalian Medical University,Dalian 116044,Chi;Department of Anesthesiology,Qingdao Municipal Hospital,Qingdao 266071,China)
出处
《国际麻醉学与复苏杂志》
CAS
2021年第3期244-247,共4页
International Journal of Anesthesiology and Resuscitation
基金
青岛市科技民生项目(19-6-1-50-nsh)。
关键词
亚低温
小鼠神经母细胞瘤细胞
氧糖剥夺/复氧
小泛素类修饰蛋白特异性蛋白酶3
Mild hypothermia
Mouse neuro-blastoma N2a cells
Oxygen glucose deprivation/reoxygenation
Small ubiquitin-like modifier proteins specific protease 3
