PINK1参与调节多巴胺的合成  

The regulatory effects of PINK1on dopamine biosynthesis

作　　者：贾焕珍[1] 鲁玲玲[1] 龚普盛[1] 付越姣[1] 赵春礼[1] 段春礼[1] 杨慧[1] 

机构地区：[1]首都医科大学神经生物学系北京神经科学研究所北京市神经再生修复研究重点实验室神经变性病教育部重点实验室,北京100069

出　　处：《基础医学与临床》2012年第1期16-20,共5页Basic and Clinical Medicine

基　　金：国家重点基础研究发展计划(2011CB504103);国家自然科学基金(30970940);北京市教育委员会科技发展计划重点项目(KZ201010025022);北京市自然基金面上项目(5102012;5102007);北京市高校人才强教资助PHR(200907113)

摘　　要：目的探讨PINK1基因对多巴胺合成的调节作用。方法用高压液相色(HPLC)电化学方法检测多巴胺(DA)含量,用Western blots和Real-Time PCR法检测DA合成限速酶酪氨酸羟化酶(TH)和多巴脱羧酶(DDC)的表达量,通过免疫共沉淀(Co-IP)和免疫组织化学染色观察PINK1和TH之间是否存在相互作用。结果 PINK1基因敲减后,细胞内DA水平为5.356±0.536μg/L,显著低于对照组的11.630±1.559μg/L(P<0.05);TH mRNA和蛋白的表达分别为0.371±0.140和0.195±0.062,显著低于对照组的1.009±0.152和0.386±0.044(P<0.05);DDC mRNA和蛋白的表达分别为0.396±0.124和0.149±0.029,显著低于对照组的1.100±0.070和0.323±0.037(P<0.05);免疫荧光化学检测PINK1和TH存在共定位,Co-IP显示PINK1和TH存在相互作用。结论 PINK1可以通过调控DA的合成酶TH和DDC的表达影响DA的合成。Objective To explore the role for PINK1 in the regulation of dopamine biosynthesis. Methods The dopaminergic MN9D cells were used as a cell model for the present study. PINK1 gene was knockdown by RNAi technique. The dopamine （DA） level, mRNA and protein expression level of tyrosine hydroxylase （TH） and dopa decarboxylase （DDC） were then detected by HPLC with electrochemical detection, real-time RT-PCR or Western blots 48h later, respectively. The association between PINK1 and TH was observed with Co-IP and immunohistochemistry. Results When PINK1 was silenced, DA content was 5.356± 0. 536 μg/L significantly lower than the control group of 11. 630 ± 1. 559μg/L （P 〈0. 05）, TH mRNA and protein expression were 0. 371 ±0. 140 and 0. 195±0. 062 significantly lower than that of control group 1. 009±0. 152 and 0. 386 ±0. 044 （P 〈0. 05）, DDC mRNA and protein expression were 0. 396 ±0. 124 and 0. 149 ±0. 029 significantly lower than that of control group 1. 009± 0. 152 and 0. 386 ± 0. 044, respectively （P 〈 0. 05）. Conclusions PINK1 has a regulatory effect on dopamine biosynthesis, through its influence on TH and DDC.

关 键 词：PINK1 RNA干扰 酪氨酸羟化酶 MN9D细胞 

分 类 号：R742.5[医药卫生—神经病学与精神病学]