锰致大鼠多巴胺能神经细胞DA分泌减少效应中PARK2基因的调控作用  

Effect of gene PARK2 in manganese-induced rat dopaminergic neurons dopamine secretion decrease

作　　者：张蓓[1] 陶冬 罗英 范希敏 范奇元[3] ZHANG Bei;TAO Dong;LUO Ying;FAN Xi-min;FAN Qi-yuan(Orthopaedics of The First People＇ s Hospital of Zunyi, Zunyi Guizhou 563009, China;Medical Administration Division of Dalian BYBO Dental Group, Dalian Liaoning 116021, China;Zunyi Medical and Pharmaceutical College, Zunyi Guizhou 563006, China;Zunyi Medical College, Zunyi Guizhou 563009, China)

机构地区：[1]遵义市第一人民医院骨科,贵州遵义563009 [2]大连拜博口腔医务部,辽宁大连116021 [3]遵义医药高等专科学校,贵州遵义563006 [4]遵义医学院,贵州遵义563099

出　　处：《毒理学杂志》2018年第2期98-101,共4页Journal of Toxicology

基　　金：国家自然科学基金(81260420);贵州省科技支撑计划项目2013-2015[黔科合SY字(2012)3140]

摘　　要：目的研究锰对经RNA沉默PARK2基因后的多巴胺(DA)能神经元细胞功能的影响。方法用原代培养新生大鼠纹状体细胞,分3类处理组,即:单纯锰处理组、慢病毒对照组和慢病毒PARK2 SiRNA干扰处理组。各处理组均用不同浓度锰(0、300和500μmol/L)处理,分别用q PCR检测PARK2表达,Western blot检测Parkin,ELISA法检测细胞分泌的DA水平。结果在各处理组中:与0μmol/L相比较300和500μmol/L染锰组PARK2表达及Parkin蛋白、DA分泌含量均下降,差异有统计学意义(P<0.05);单纯染锰处理组及慢病毒对照组与PARK2 SiRNA干扰处理组比较,相同染锰剂量的PARK2表达及Parkin蛋白、DA分泌含量均降低,差异有统计学意义(P<0.05);单纯染锰处理组与慢病毒对照组比较,相同染锰剂量的PARK2表达及Parkin蛋白、DA分泌含量均无明显变化,差异无统计学意义(P>0.05)。结论对大鼠多巴胺(DA)能神经元细胞进行PARK2RNA干扰后再染锰,可加重其DA分泌量的降低,锰暴露作用下DA能神经元的功能与PARK2的改变相关。Objective To study the effect of manganese on dopaminergic neurons after RNA silenced-PARK2 gene. Methods Primary cultured neonatal rat striatal neurons were divided into three groups： manganese treatment group(Mn group） 、lentivirus control group(sh-C group） and PARK2 SiRNA interfere group(sh group）. Cells were exposed to manganese at different concentrations(0,300 and 500 μmol/L）.PARK2 mRNA expression,protein Parkin and dopamine level were tested by using Real-time PCR,Western blot and Elisa kit respectively.Results Each group neurons PARK2 mRNA,Parkin and dopamine level after manganese exposure(300 and 500 μmol/L） were decreased compared with the 0 μmol/L manganese exposure group(P 〈0. 05）; Neurons PARK2 mRNA expression,Parkin and dopamine level were increased in Mn and sh-C group compared with the sh group at the same manganese concentration(P〈 0. 05）. There were no sigmicant difference at PARK2 mRNA,Parkin and dopamine level in Mn and sh-C group at the same manganese concentration(P〉 0. 05）.Conclusion Manganese deteriorated the dopamine decreasing of the PARK2 SiRNA interfered rat dopaminergic neurons,dopaminergic neurons function were involved with PARK2 level after manganese exposure.

关 键 词：锰 多巴胺 PARK2 神经细胞 

分 类 号：R114[医药卫生—卫生毒理学]