纳米氧化锌对SH-SY5Y神经细胞毒性损伤作用及分子机制研究  

Toxicity and molecular mechanisms of ZnO nano-particles on SH-SY5Y cells

作　　者：万彬彬[1] 何浩[1] 江维 赵宇侠[3] 金京娥 金惠[5] 

机构地区：[1]武汉市第一医院风湿免疫科,湖北武汉430022 [2]马应龙药业集团股份有限公司,湖北武汉430064 [3]淮海工学院化学工程学院,江苏连云港222005 [4]武汉市新洲区中医院,湖北武汉430400 [5]同济大学生命科学与技术学院,上海200092

出　　处：《中国药理学通报》2013年第11期1587-1591,共5页Chinese Pharmacological Bulletin

基　　金：中国博士后基金(No 2013531221);连云港市科技局农业攻关项目(No NY1007)

摘　　要：目的研究纳米氧化锌(ZnO NPs)对SH-SY5Y人神经细胞的毒性作用及其可能的分子机制。方法 MTT法检测ZnO NPs对SH-SY5Y细胞存活率的影响;TUNEL法检测ZnO NPs对SH-SY5Y细胞的形态学影响;流式细胞术观察ZnO NPs对SH-SY5Y细胞周期的影响及凋亡诱导作用;Real-time PCR检测凋亡相关基因Bcl-2、Bax、Survivin的mRNA水平变化;Western blot测定Bcl-2、Bax、Survivin及Cyto-C的蛋白表达并进行光密度定量。结果 ZnO NPs显示出对SHSY5Y的细胞毒性,呈现浓度依赖性;15 mg·L-1ZnO NPs作用后,TUNEL检测呈阳性反应;与未处理组相比,SH-SY5Y细胞中细胞周期阻滞于G1期并产生凋亡(P<0.05);抗凋亡基因Bcl-2、Survivin的mRNA水平和蛋白水平下调(P<0.05),促凋亡蛋白Bax的mRNA水平和蛋白水平上调,同时细胞Cyto-C的蛋白表达上调(P<0.05)。结论 ZnO NP下调SH-SY5Y的细胞存活率,诱导细胞周期阻滞和凋亡产生,显示出对神经细胞的毒性损伤作用,其机制与激活线粒体凋亡通路及抑制Survivin的转录和表达有关。Aim To examine the toxic effects of ZnO NPs on neuroblastoma cell line SH-SY5Y and explore the molecular mechanisms underlying the effects. Methods The effects of ZnO NPs on the cell viability were detected by MTT assays and the effects on the cell cycle arrest and apoptotic induction were examined by flow cytometry. The regulations on mRNA level of Bcl- 2, Bax and Survivin were measured by Real-time PCR; the protein expressions of Bcl-2, Bax, Survivin and Cyto-C were detected by Western blot and measured by densitometry. Results ZnO NPs exhibited eytotoxicity in SH-SY5Y cells dose-dependently. TUNEL results suggested ZnO NPs（ 15 mg· L ^-1） in-duced apoptotic characteristics in SH-SY5Y cells. ZnO NPs induced G1 phase cell cycle arrest and apoptosis （P 〈 0.05 ）. The mRNA level and protein expression of Bax were upregulated whereas the protein expression of Bcl-2, Survivin were downregulated （ P 〈 0.05 ）. Conclusion ZnO NPs exhibit toxicity in neuroblastoma SH-SY5Y cells and induce cell cycle arrest and apoptosis in SH-SY5Y cells. The molecular mechanisms are associated to regulations on Bcl-2,Survivin and Bax and triggering mitochondrial apoptotie pathway.

关 键 词：纳米氧化锌 神经毒性 细胞周期阻滞 线粒体凋亡 Bcl-2 Bax Survivin 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]