NMNAT1对MES23.5帕金森病细胞模型Caspase8及TH蛋白表达影响  

EFFECT OF NMNAT1 ON EXPRESSIONS OF CASPASE8 AND TH IN CELL MODEL OF PARKINSON'S DISEASE MES23.5 CELLS

作　　者：刘琳[1] 刘洪欣[1] 李春燕[1] 谢安木[1] 

机构地区：[1]青岛大学附属医院神经内科,山东青岛266003

出　　处：《齐鲁医学杂志》2016年第6期663-666,共4页Medical Journal of Qilu

基　　金：山东省自然科学基金资助项目(ZR2012CM028)

摘　　要：目的探讨烟酰胺单核苷酸腺苷酰转移酶1(NMNAT1)对帕金森病(PD)细胞模型MES23.5细胞Caspase8、酪氨酸羟化酶(TH)蛋白表达的影响。方法将MES23.5细胞分为6组,pEGFP组以NMNAT1高表达(NMNAT1-OE)空载质粒转染MES23.5细胞;pEGFP+MPP^+组以NMNAT1-OE空载质粒转染MES23.5细胞后,再向细胞内加入1-甲基-4-苯吡啶离子(MPP^+)100mmol/L;NMNAT1-OE+MPP^+组以NMNAT1-OE质粒转染MES23.5细胞,再向细胞内加入MPP^+100 mmol/L;pMagic4.1组以NMNAT1RNAi空载质粒转染MES23.5细胞;pMagic4.1+MPP^+组以NMNAT1 RNAi空载质粒转染MES23.5细胞,再加入100 mmol/L的MPP^+;NMNAT1-RNAi+MPP^+组以NMNAT1RNAi质粒转染细胞后,再加入100mmol/L的MPP^+。应用Western印迹分析方法检测各组TH、Caspase8蛋白表达。结果pEGFP+MPP^+组、pEGFP组、NMNAT1-OE+MPP^+组TH表达依次升高,Caspase8表达依次降低,3组间比较差异有显著性(F=18.46、13.26,P<0.05)。pMagic4.1组、pMagic4.1+MPP^+组、NMNAT1-RNAi+MPP^+组TH表达依次降低,差异有显著性(F=11.78,P<0.05);pMagic4.1+MPP^+组Caspase8表达最高,NMNAT1-RNAi+MPP^+组次之,pMagic4.1组最低,差异有显著意义(F=17.65,P<0.05)。结论 NMNAT1可能通过抑制Caspase8的表达、同时增加TH的表达而发挥神经保护作用。Objective To investigate the effect of nicotinamide mononucleotide adenylyltransferase （NMNAT1） on the expressions of Caspase8 and TH （Tyrosine hydroxylase） in cell model of Parkinson’s disease （PD） MES23.5 cells. Methods The MES23.5 cells were divided into six groups as： pEGFP group-the cells were transfected with high expression NMNAT1-OE empty plasmids; pEGFP＋MPP^＋group-the cells in pEGFP group were treated with 1-methyl-4-phenylpyridinium （MPP^＋; 100 mmol/L）; NMNAT1-OE＋MPP^＋ group-MES23.5 cells were transfected with NMNAT1-OE plasmids and followed with MPP^＋100 mmol/L; pMagic4.1 group-transfected with NMNAT1 RNAi vehicle plasmids into MES23.5 cells; pMagic4.1＋MPP＋group-MES23.5 cells were transfected with NMNAT1 RNAi vehicle plasmids and 100 mmol/L of MPP^＋ added; NMNAT1-RNAi＋MPP^＋ group-NMNAT1 RNAi plasmids were transfected into MES23.5 cells, and then treated with MPP^＋. The expressions of Caspase8 and TH in the six groups were detected using Western blots. Results The expression of TH in pEGFP, pEGFP＋MPP^＋ and NMNAT1-OE＋ MPP^＋groups was increased in sequence, and that of Caspase8 decreased, the differences among the three groups were significant （F=18.46,13.26;P〈0.05）. The expression of TH in pMagic4.1, pMagic4.1＋MPP^＋, and NMNAT1-RNAi＋MPP^＋ groups was decreased in turn, and the expression of Caspase8 in the pMagic4.1＋MPP^＋ group was the highest, followed by NMNAT1-RNAi＋MPP^＋group and the lowest was noted in the pMagic4.1 group, the difference was significant （F=17.65,P〈0.05）. Conclusion Our results suggest that NMNAT1 may provide a potential neuroprotective effect by down-regulating the expression of Caspase8 and up-regulating the expression of TH.

关 键 词：烟酰胺单核苷酸腺苷酰转移酶1 帕金森病 半胱氨酸天冬氨酸蛋白酶8 酪氨酸单氧化酶 

分 类 号：R338.2[医药卫生—人体生理学]