中文摘要：

目的探讨帕金森病相关基因PINK1对酪氨酸羟化酶（tyrosine hydroxylase,TH）的表达调控作用。方法分别建立PINK1基因敲减组（转染携带有PINK1小干扰RNA片段的质粒）和PINK1基因过表达（转染PINK1真核表达质粒载体）的MN9D细胞模型,通过实时定量PCR法（real-time RT-PCR）和Western blot法分别检测THmRNA水平和蛋白水平,以评测改变细胞内PINK1含量对于TH表达的影响。结果在转染后48 h检测时发现,PINK1基因敲减组较对照组TH mRNA水平下调可达76.3%（P〈0.001）;与对照组相比,蛋白水平亦明显降低,降低幅度达74.1%（P〈0.001）。与此同时,检测到p-TH的蛋白水平也降低约75.1%（P〈0.001）,但是p-TH/TH的水平没有明显改变。在MN9D细胞中瞬时转染PINK1野生型及突变体G309D,发现过表达野生型PINK1组TH蛋白水平明显增加（增加115.9%,P〈0.05）,但是过表达PINK1突变体G309D组无明显差异（P〉0.05）。结论 PINK1可调节多巴胺合成过程中的限速酶TH的转录及蛋白的表达,而其G309D突变体表现为此项功能的缺失。

英文摘要：

Objective To investigate the regulatory effects of PINK1 on tyrosine hydroxylase（ TH） gene expression and its mechanism.Methods The MN9 D cells were divided into four groups： PINK1 knocking-down group（ transfected with small interference RNA for PINK1）and its control group（ transfected with scramble sequence）,PINK1 overexpressing group（ transfected with PINK1 expressing plasmid） and its control group（ transfected with empty plasmid,vector group）. Then mRNA and protein levels of TH were detected by real-time PCR and Western blot,respectively. Results Compared with control group,TH mRNA level decreased by 76. 3% after transfection for 48 h in PINK1 knocking-down group（ P 0. 001）,the protein expression level decreased by 74. 1%（ P 0. 001）,and p-TH protein level also decreased by 75. 1%（ P 0. 001）,but p-TH / TH showed no obvious changes. Meanwhile,compared with vector group,TH protein levels increased by 115. 9% in PINK1-WT overexpressing group（ P 0. 05）,but no significant change in PINK1-G309 D overexpressing group（ P 0. 05）. Conclusion These data suggest that PINK1 may play a role in the regulation of TH expression.