烟酰胺单核苷酸腺苷酰转移酶1对MES23.5帕金森病细胞模型过氧化氢酶及酪氨酸羟化酶蛋白表达的影响

Effect of NMNAT1 on Expression of Catalase and Tyrosine Hydroxylase in Cell Models of Parkinson's Disease MES23.5

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(MES 23.5细胞)过氧化氢酶(CAT)、酪氨酸羟化酶(TH)表达的影响。方法利用质粒转染或SiRNA技术分别处理MES 23.5细胞,依据不同处理方式分为6组:pEGFP组、pEGFP+MPP+组、NMNAT1-OE+MPP+组、pMagic4.1组、pMagic4.1+MPP+组和NMNAT1-RNAi+MPP+组。采用Western blot法检测各组MES23.5细胞的CAT和TH半定量表达水平。结果 NMNAT1呈高表达的NMNAT1-OE+MPP+组、pMagic4.1+MPP+组CAT表达量和TH蛋白表达量均升高;NMNAT1呈低表达的pEGFP+MPP+组、NMNAT1-RNAi+MPP+组CAT蛋白表达也有所降低,TH蛋白表达量下降。结论 NMNAT1通过增加CAT表达水平参与抗氧化应激,并且增加TH表达,具有一定的神经保护作用。

Aim To investigate the effects ofnicotinamide mononucleotide adenylyltransferase （NMNAT1） on the expression of catalase （CAT） and tyrosine hydroxylase （TH） in cell Models of Parkinson＇s disease （PD） MES23.5 cells. Methods Before parkinsonian cell model establishment, synthetic NMNAT1 overexpression （NMNAT1-OE） vehicle plasmids, NMNAT1 overexpression plasmids, NMNAT1 siRNA vehicle plasmids and NMNAT1 siRNA （NMNAT1 RNAi） plasmids were transfected into MES23.5 cells using cell transfection techniques. Cells were divided into 6 groups： pEGFP group, pEGFP＋MPP＋ group, NMNAT1-OE＋MPP＋ group, pMagic4.1 group, pMagic4. I＋MPP group, NMNAT1-RNAi＋MPP＋ group. The protein levels of caspase 8 and TH in the 6 groups were measured by Western blot. Results Compared with pEGFP group, marked decreased in the protein level of CAT was observed in pEGFP＋MPP＋ group （t=9.01, P〈0.001）, and TH expression decreased （t=0.32, P=0.012）. Compared with pEGFP＋MPP＋group, CAT expression increased （t=2.51, P= 0.033） and TH expression increased （t=0.45, P=0.002） in NMNAT1-OE＋MPP＋ group. Compared with pMagic4.1 control group, marked decreased in the protein level of CAT （t=2.47, P=0.036） and decrease in the protein level of TH （t=0.49, P=-0.001） was observed in pMagic4.1 ＋MPP＋ group. Compared with pMagic4. I＋ MPP＋ group, CAT expression decreased （t=3.53, P=0.006） and TH expression decreased （t=0.39, P=0.004） in NMNAT1-RNAi＋MPP＋ group. Conclusion The results suggested that NMNAT1 may be effective to up-regulate the expression of CAT which is one of antioxidant enzyme against oxidative injury and TH. NMNAT1 may play an important role in protecting DA neurons through CAT.