摘要
目的:研究多巴胺(DA)对多巴胺能细胞的毒性作用、NAD(P)H:醌氧化还原酶1(NQO1)的保护作用及其机制。方法:运用MTT法检测多巴胺对神经母细胞瘤细胞(SH-SY5Y)活性的影响;运用脂质体转染或Ⅱ相酶诱导剂预处理的方法对细胞进行预处理,并用免疫荧光方法检测转染效率;运用蛋白质印迹(Western blotting)方法检测细胞经过不同处理后胞内NQO1蛋白的表达情况;运用醌蛋白检测方法(硝基四氮唑蓝/甘氨酸法)检测细胞经不同处理后胞内醌化蛋白含量的变化。结果:多巴胺对细胞具有剂量依赖性毒性,且与细胞内醌化蛋白的含量相关;脂质体转染和Ⅱ相酶诱导剂萝卜硫素(SF)均能使细胞内NQO1表达量增高;NQO1高表达能缓解多巴胺对细胞造成的毒性;细胞内NQO1表达量增高能减少多巴胺引起的细胞内醌化蛋白含量。结论:细胞内NQO1的过表达可以减轻多巴胺对细胞产生的毒性作用。
AIM：To determine the influence of NAD（P）H：quinone oxidoreductase 1（NQO1） on dopamine-induced toxicity in dopaminergic cells.METHODS： MTT assay was used to determine the toxic curve of dopamine in SH-SY5Y cells.Lipofection was applied to transfect SH-SY5Y cells with an NQO1 expression plasmid.The endogenous and transfected NQO1 expression was detected by immunofluorescence staining and Western blotting.The content of cellular quinone protein was measured by nitroblue tetrazolium（NBT） method.RESULTS： Dopamine reduced SH-SY5Y cell proliferation in a dose-dependent manner,which was correlated with an increase in the content of quinone protein.Increased expression of NQO1 by transient transfection or by phase II enzyme inducer sulforaphane treatment alleviated dopamine-induced toxicity and reduced the content of cellular quinone protein.CONCLUSION： Increased NQO1 expression protects SH-SY5Y cells against cytotoxicity caused by dopamine.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2011年第10期1917-1921,共5页
Chinese Journal of Pathophysiology
基金
教育部985人才基金资助项目(No.50001477)
