TAp63对MPP^(+)处理的SH-SY5Y细胞半胱氨酸天冬氨酸蛋白酶-9和酪氨酸羟化酶表达的影响  

Effect of TAp63 Gene on Caspase-9 and Tyrosine Hydroxylase Expression in MPP^(+)Induced Parkinson’s Cell

作　　者：刘晨[1,2] 任继文 谢安木 LIU Chen;REN Ji-wen;XIE Aan-mu(Department of Neurology,the Affiliated Hospital of Qingdao University,Qingdao 266003,China;Qingdao Emergency Center,Qingdao 266000,China;Institute of Cerebrovascular Diseases,Qingdao University,Qingdao 266000,China)

机构地区：[1]青岛大学附属医院神经内科,青岛266003 [2]青岛市急救中心,青岛266000 [3]青岛大学脑血管病研究所,青岛266000

出　　处：《中国临床神经科学》2023年第2期132-137,共6页Chinese Journal of Clinical Neurosciences

基　　金：国家自然科学基金资助项目(编号:81971192)。

摘　　要：目的探讨TAp63基因表达水平对1-甲基-4-苯基吡啶离子(MPP^(+))诱导的帕金森病(PD)细胞模型细胞活性、酪氨酸羟化酶(TH)和凋亡因子半胱氨酸天冬氨酸蛋白酶-9(caspase-9)的影响。方法将小鼠神经母细胞瘤细胞(SH-SY5Y细胞系)依据不同处理方法分为6组:(1)对照组,无任何干预措施;(2)MPP^(+)处理组,MPP^(+)10μmol·L^(-1)处理24 h;(3)TAp63-NC组,用携带无义序列的慢病毒颗粒转染48 h;(4)TAp63-NC+MPP^(+)组,用携带无义序列的慢病毒颗粒转染48 h后,加MPP^(+)10μmol·L^(-1)处理24 h;(5)TAp63过表达组,用携带过表达TAp63基因的慢病毒颗粒转染48 h;(6)TAp63过表达+MPP^(+)组,用携带过表达TAp63基因的慢病毒颗粒转染48 h,换液后予以MPP^(+)10μmol·L^(-1)处理24 h。采用CCK-8法检测6组细胞的存活率;Western blot法检测TAp63、TH、caspase-9表达水平。结果CCK-8检测显示,与对照组比较,MPP^(+)组细胞活性显著下降(P<0.01)。与TAp63-NC+MPP^(+)组比较,TAp63过表达+MPP^(+)组细胞存活率升高,差异有显著统计学意义(P<0.001)。Western blot检测显示:与对照组比较,MPP^(+)组的TAp63蛋白和TH蛋白表达量降低,而caspase-9表达量升高(均P<0.05);与TAp63-NC+MPP^(+)组比较,TAp63过表达+MPP^(+)组TAp63蛋白和TH蛋白表达量升高,而caspase-9蛋白表达水平降低(均P<0.05)。结论TAp63基因过表达可提升MPP^(+)诱导的PD细胞的存活率,提高TH表达量,降低caspase-9表达水平,进一步发挥其潜在的神经保护作用。Aim To investigate the effect of TAp63 gene expression level on cell viability,tyrosine hydroxylase(TH)and caspase-9 expression level in Parkinson’s cell model induced by 1-methyl-4-phenyl-pyridinium(MPP^(+)).Method Mouse neuroblastoma cells(SH-SY5Y cell line)were divided into 6 groups according to different treatment methods:①Control group,without any intervention.②MPP^(+)treatment group,SH-SY5Y cells treated with 10μmol·L^(-1)MPP^(+)for 24 hours.③TAp63-NC treatment group,SH-SY5Y cells treated with lentivirus particles with nonsense sequence for 48 hours.④TAp63-NC+MPP^(+)treatment group,SH-SY5Y cells treated with lentivirus particles with nonsense sequence for 48 hours,and then treated with 10μmol·L^(-1)MPP^(+)for 24 hours.⑤TAp63 overexpression treatment group,lentivirus particles carrying TAp63 overexpression gene continuously transfected for 48 hours.⑥TAp63 overexpression+MPP^(+)treatment group,lentivirus particles carrying TAp63 overexpression gene continuously transfected for 48 hours,and 10μmol·L^(-1)MPP^(+)for 24 hours after changing the solution.CCK-8 assay was used to test the cell viability.Western blot was used to detect the expression levels of TAp63,TH and casepase-9.Results Compared with the control groups,cell survival rates in MPP^(+)were decreased(P<0.01)in CCK-8 assay tests.Besides,compared with TAP63-NC+MPP^(+)group,cell livability in TAP63-NC+MPP^(+)group were increased,the difference was statistically significant(P<0.001).Compared with the control group,TAp63 protein expression and TH expression in MPP^(+)group were decreased,while caspase-9 expression was increased(all P<0.05)in Western blot analysis.Compared with TAP63-NC+MPP^(+)group,TAp63 protein expression level and TH expression level were increased in TAp63 overexpression+MPP^(+)group,while caspase-9 protein expression level was decreased(all P<0.05).Conclusion Overexpression of TAp63 gene can improve MPP^(+)cell viability,increase the expression level of TH and decrease the expression level of caspase-9 in Parkinso

关 键 词：帕金森病 TAp63基因 酪氨酸羟化酶 半胱氨酸天冬氨酸蛋白酶-9 SH-SY5Y细胞系 1-甲基-4-苯基吡啶离子 

分 类 号：R742.5[医药卫生—神经病学与精神病学] Q189[医药卫生—临床医学]