TAp63基因对帕金森病细胞模型中凋亡因子表达的影响  

Effect of TAp63 Gene on the Expression of Apoptotic Factors in MPP+Induced Parkinson’s Cell Model

作　　者：石万达 刘贤[1] 窦凯昕 刘玉梅[1] 谢安木[1] SHI Wan-da;LIU Xian;DOU Kai-xin;LIU Yu-mei;XIE An-mu(Department of Neurology,Affiliated Hospital of Qingdao University,Qingdao 266003,China)

机构地区：[1]青岛大学附属医院神经内科,青岛266003

出　　处：《中国临床神经科学》2022年第3期241-246,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^(+))诱导的帕金森病细胞模型中凋亡因子B细胞淋巴瘤-2(Bcl-2)蛋白、Bcl-2相关x(Bax)蛋白、髓样细胞白血病-1(Mcl-1)蛋白表达水平的影响。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4组:(1)对照组,无任何干预措施;(2)MPP^(+)处理组,添加10μmol·L^(-1)MPP^(+)处理24 h;(3)TAp63-NC+MPP^(+)组,用携带无义序列的慢病毒颗粒转染48 h后,加10μmol·L^(-1)MPP^(+)处理24 h;(4)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10μmol·L^(-1)MPP^(+)处理24 h。采用Western blot法检测4组TAp63、Bcl-2、Bax、Mcl-1凋亡因子的表达水平,流式细胞术检测4组凋亡细胞数。结果Western blot检测显示:与对照组比较,MPP^(+)处理组TAp63蛋白表达降低(P<0.01);与TAp63-NC+MPP^(+)组比较,TAp63过表达+MPP^(+)组TAp63蛋白表达水平升高(P<0.01)。与对照组比较,MPP^(+)处理组Bax表达水平升高(P<0.01);Bcl-2及Mcl-1表达水平降低(均P<0.01)。与TAp63-NC+MPP^(+)组比较,TAp63过表达+MPP^(+)组Bax表达水平降低(P<0.01);Bcl-2和Mcl-1表达水平升高(分别P<0.05和P<0.01)。流式细胞术提示:与对照组比较,MPP^(+)处理组凋亡细胞数增加(P<0.01);与TAp63-NC+MPP^(+)组比较,TAp63过表达+MPP^(+)组凋亡细胞数显著减少(P<0.01)。结论TAp63基因过表达可改变帕金森病细胞模型中促凋亡因子Bax、Bcl-2和Mcl-1表达水平,发挥潜在的神经保护作用。Aim To investigate the effect of TAp63 gene expression on the expression of B cell lymphoma-2 protein(Bcl-2),Bcl-2 associated x protein(Bax)and myeloid leukemia-1 protein(Mcl-1)apoptosis factors in Parkinson’s model induced by 1-methyl-4-phenyl-pyridinium(MPP^(+)).Method Mouse neuroblastoma cells(SH-SY5Y cell line)were divided into 4 groups according to different treatment methods:(1)Control group:without any intervention.(2)MPP^(+) treatment group:SH-SY5Y cells were treated with 10μmol·L^(-1)MPP^(+) for 24 hours.(3)In TAp63-NC+MPP^(+) treatment group,SH-SY5Y cells were treated with lentivirus particles with nonsense sequence for 48 hours,and then treated with 10μmol·L^(-1)MPP^(+) for 24 hours.(4)In the TAp63 overexpression+MPP^(+) treatment group,lentiviral particles carrying TAp63 overexpression gene were continuously transfected for 48 hours,and 10μmol·L^(-1)MPP^(+)for 24 hours after changing the solution.The expression levels of TAp63,Bcl-2,Bax and Mcl-1 apoptosis factors in the four groups of cells were detected by Western blot.The number of apoptotic cells in the four groups was dectected by flow cytometry.Results The results of Western blot showed that the expression of TAp63 protein in MPP^(+)treated group was lower than that in the control group,and that in TAp63 overexpression+MPP^(+)treated group was higher than that in TAp63-NC+MPP^(+)group(P<0.01).Compared with the control group,the expression of Bax in MPP^(+)treated group increased,while the expression of Bcl-2 and Mcl-1 decreased(all P<0.01).Compared with TAp63-NC+MPP^(+)group,the expression of Bax decreased,while the expression of Bcl-2 and Mcl-1 increased in TAp63 overexpression+MPP^(+)group(P<0.05,0.01,respectively).Flow cytometry showed that the number of apoptotic cells in MPP^(+)treated group was higher than that in the control group,and that in TAp63 overexpression+MPP^(+)treated group was significantly lower than that in TAp63-NC+MPP^(+)group(P<0.01).Conclusion Overexpression of TAp 63 gene can reduce the expression of pro-apoptoti

关 键 词：TAp63基因 帕金森病 SH-SY5Y细胞系 凋亡因子 1-甲基-4-苯基吡啶离子 

分 类 号：Q186[生物学—普通生物学] R742.5[医药卫生—神经病学与精神病学]