丙烯酰胺对NB-1细胞钙稳态的影响  

Effect of acrylamide exposure on calcium homeostasis in NB-1 cells

作　　者：李冲[1] 王茜[1] 曹燕花[1] 刘楠[1] 关维俊[1] 王学生[1] 肖经纬[2] 李斌[2] 李忠生[2] 

机构地区：[1]河北联合大学公共卫生学院,河北唐山063000 [2]中国疾病预防控制中心职业卫生与中毒控制所,北京100050

出　　处：《中国工业医学杂志》2012年第3期194-196,共3页Chinese Journal of Industrial Medicine

基　　金：国家自然基金青年基金(30901219)

摘　　要：目的研究丙烯酰胺(AM)对神经细胞钙稳态的影响,探讨AM致神经损伤的可能机制。方法将人神经母细胞瘤(NB-1)细胞诱导分化成熟后,以不同浓度AM进行体外染毒,采用噻唑蓝(MTT)比色法检测神经元损伤程度,并应用ATP酶检测试剂盒测定不同损伤程度的细胞Ca2+-ATP酶和Na+-K+-ATP酶活力的变化。以Fluo-3/AM为荧光指示剂,用激光共聚焦方法检测细胞内游离钙离子浓度的瞬时变化情况。结果 AM染毒组成熟NB-1细胞存活率有明显下降,提示AM对神经细胞有毒性作用;AM染毒组Ca2+-ATP酶和Na+-K+-ATP酶活力值也均下降,差异具有统计学意义(P<0.05);AM作用于成熟NB-1细胞可立即引起细胞内钙离子浓度升高。结论 AM可能通过影响神经细胞钙稳态产生毒性作用。Objective To observe the effect of acrylamide （AM） on calcium homeostasis in mature neurons for exploring the possible mechanisms of nerotoxicity by acrylamide. Methods Human neuroblastoma cells （ NB-1 ） induced to differentiated mature neurons with db-cAMP were exposed to different concentration of AM, then, detect their cell death rate with thiazolyl blue exlusion-MTY assay, and the activities of Ca2 ＋ -ATPase and Na ＋ -K ＋ -ATPase by ATPase kits. Meantime, laser confocal scanning microscope was used for intracellular calcium level measurement. Results The results showed that the survival rate, the activities of Ca2 ＋ -ATPase and Na ＋ -K ＋ -ATPase were all obviously decreased in AM exposed groups （ P 〈 0.05 ） ; meanwhile, 30 minutes exposure to AM might induce a rapid rise of intracellular calcium. Conclusion The results suggested that AM has obvious neurotoxicity, the disturbance of calcium homeostasis may involved in the mechanism of its neurotoxicity.

关 键 词：丙烯酰胺(AM) NB-1 db-cAMP CA2+-ATP酶 NA+-K+-ATP酶 

分 类 号：R994[医药卫生—毒理学]