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目的研究正己烷代谢产物2,5-己二酮对运动神经元钙稳态的影响,探讨正己烷中毒致周围神经损伤的机制。方法对运动神经元细胞系VSC4．1细胞不同浓度的2,5-己二酮体外染毒,用四甲基偶氮唑蓝(MTT)法检测细胞活性的变化;同时检测Ca2+-Mg2+ATP酶和Na+-K+-ATP酶活力。以Fluo-3／AM为荧光指示剂,激光共聚焦方法检测细胞内游离钙浓度的瞬时变化状况;用流式细胞仪检测细胞内钙平均浓度的变化。结果经2．5、5．0、7．5、10．0、15．0、20．0 mmol／L浓度的2,5-己二酮对VSC4．1细胞染毒后与对照组比较,Ca22+-Mg2+ATP酶和Na+-K+-ATP酶的活力均下降。33．5 mmol／L的2,5-己二酮可在10 s内引起VSC4．1运动神经元细胞内钙浓度升高,40 s后恢复到基线水平。VSC细胞与2,5-己二酮接触后即刻引起细胞内钙水平的增加,10 min后达到最高。结论2,5-己二酮可能通过干扰细胞钙稳态产生神经毒性。
Objective To investigate the effect of 2,5-hexanedione, a metabolite of n-hexane, on calcium homeostasis in motor neurons and to explore the mechanism of peripheral nerve injury caused by n-hexane poisoning. Methods Different concentrations of 2,5-hexanedione were in vitro exposed to different concentrations of VSC4.1 cells. MTT assay was used to detect the changes of cell viability. Simultaneous detection of Ca2 + -Mg2 + ATP Enzyme and Na + -K + -ATP enzyme activity. Fluo-3 / AM was used as a fluorescence indicator to detect the transient changes of intracellular free calcium concentration by confocal laser scanning microscope. The change of intracellular calcium concentration was detected by flow cytometry. Results After VSMC4.1 cells were treated with 2,5-hexanedione at concentrations of 2.5, 5.0, 7.5, 10.0, 15.0 and 20.0 mmol / L, compared with the control group, The activities of Ca22 + -Mg2 + ATPase and Na + -K + -ATPase decreased. 33.5 mmol / L of 2,5-hexanedione induced intracellular Ca2 + concentration in VSC4.1 motor neurons within 10 s, and returned to baseline level after 40 s. Immediately after exposure of VSC cells to 2,5-hexanedione, intracellular calcium levels increased, reaching the highest level after 10 min. Conclusion 2,5-Hexanedione may produce neurotoxicity by interfering with the calcium homeostasis.