1，2－二氯乙烷对大鼠神经细胞内钙离子浓度的影响

Change of Ca^(2+) Concentration of Rat Neuron Induced by 1,2-dichloroethane

[目的]用体外实验方法研究1,2-二氯乙烷(1,2-DCE)染毒后大鼠神经细胞内钙离子(Ca2+)浓度的变化。[方法]体外培养新生SD大乳鼠脑皮质细胞,分别用0.5、1.0、2.0mL/L 1,2-DCE染毒,另设对照组,观察各组神经细胞形态学变化。同时应用激光共聚焦显微镜测定各组细胞内Ca2+浓度,探讨1,2-DCE对大鼠神经细胞内Ca2+浓度的影响。[结果]神经细胞染毒后,神经细胞形态发生明显改变:胞体肿胀崩解、胞核模糊不清、突触变短变粗、细胞间连接减少、细胞膜不完整;随着染毒剂量的增高,神经细胞损伤的严重程度呈上升趋势。各染毒组神经细胞活力较对照组有所下降,差异有统计学意义(P<0.01);染毒2h后随着染毒剂量的增加,各组神经细胞内Ca2+浓度呈上升趋势,各染毒组细胞与对照组比较差异有统计学意义(P<0.01);染毒24h后,低、中剂量组与对照组细胞内Ca2+浓度比较差异有统计学意义(P<0.01),高剂量组与对照组比较差异无统计学意义(P>0.05)。[结论]1,2-DCE可以导致神经细胞水肿坏死,有剂量依赖关系,神经细胞内Ca2+浓度亦见升高,提示Ca2+可能与1,2-DCE致神经细胞中毒性脑水肿有关。

[ Objective ] To study the change of intracellular calcium concentration （ cell [ Ca^2＋ ] i ） in rat neuron induced by 1,2-dichloroethane （ 1,2-DCE ） in vitro. [ Methods ] Neurocytes of new born SD rats were cultured in vitro, which were administrated with different doses of 1,2-DCE, including control group, low dose group（ 0.5 mL/L ）, middle dose group（ 1.0mL/L ） and high dose group（ 2.0mL/L ）. The cell structural morphological changes were observed under light microscope, and proliferation of cells was detected by Cell Counting Kit-Ⅷ. The cell[ Ca^2＋ ]i was determined by laser confocal microscopy. [ Results ] 1,2-DCE can injure the normal structural morphology of neurocyte： the cell body swelled and brokedown, the karyon slurred or disappeared, the axone shortened and became thick, the connection among neurocytes was reduced, the cell membrane was half-baked, the injury of neurocytes became severer with the dose increase of the 1,2-DCE. Proliferation of neurocytes 1,2-DCE groups＇ was lower than control group＇s, with significant statistically difference （ P〈0.01 ）. The cell [ Ca^2＋ ] i increased with the dose increasing of the 1,2-DCE in 2 hours after 1,2-DCE administration, there was statistically significant difference between 1,2-DCE groups and control group（ P〈0.01 ）; but the cell[ Ca^2＋ ]i decreased in 24hours after 1,2-DCE administration, and the[ Ca^2＋ ]i in the high dose group was lower than those in the low and middle dose group＇s. There was no statistically significant difference of the [ Ca^2＋ ] i between the high dose group and the control group （P 〉 0.05 ）, but there were significant differences between low dose group, middle dose group and control group （P 〈 0.01 ）. [ Conclusion ] 1,2-DCE can damage the normal structural morphology of neurocytes, and became severer with the dose increase of the 1,2-DCE, and can increase [ Ca^2＋ ] i after administered with 1,2-DCE. Thus Ca^2＋ may play an important role in acute intoxicated encephalopathy induced by 1,2-dichloroethane.