丙烯酰胺对NB-1细胞钙稳态的影响

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。

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

Objective To observe the effect of acrylamide(AM)on calcium homeostasis in mature neurons for exploring the pos sible mechanisms of nerotoxicity by acry lamide.Methods Human neuroblastoma cells(NB-1)induced to differentiated mature n eurons with db-cAMP were exposed to diff erent concentration of AM,then,detect th eir cell death rate with thiazolyl blue exlusion—MTT assay,and the activities of Ca2+-ATPase and Na+-K+-ATPase by A TPase kits.Meantime,laser confocal scann ing microscope was used for intracellula r calcium level measurement.Results The results showed that the survival rate,th e activities of Ca2+-ATPase and Na+K+-ATPase were all obviously decreased in AM exposed groups(P<0.05);meanwhile,3 0 minutes exposure to AM might induce a rapid rise of intracellular calcium.Conclusion The results suggested that AM has obvious neurotoxicity,the disturbance o f calcium homeostasis may involved in th e mechanism of its neurotoxicity.