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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 investigate the effect of acrylamide (AM) on the calcium homeostasis in neurons and to explore the possible mechanism of AM injury. Methods Human neuroblastoma (NB-1) cells were induced to differentiate into mature cells and then in vitro treated with different concentrations of AM. MTT assay was used to detect the neuronal damage. The level of neuronal damage was measured by ATPase assay kit Changes of cell Ca2 + -ATPase and Na + -K + -ATPase activity in different degree of injury. With Fluo-3 / AM as fluorescent indicator, the laser confocal method was used to detect the transient changes of intracellular free calcium concentration. Results The survival rate of mature NB-1 cells in AM-treated group was significantly decreased, suggesting that AM had a toxic effect on nerve cells. The activity of Ca2 + -ATPase and Na + -K + -ATPase in AM-treated group also decreased, with statistically significant difference Significance (P <0.05); AM effect on mature NB-1 cells can immediately lead to increased intracellular calcium concentration. Conclusion AM may exert toxic effects on the calcium homeostasis of neurons.