MDMA对原代培养乳鼠脑皮质神经元的损伤机制

目的探讨3,4-亚甲双氧甲基安非他明(MDMA)对原代培养乳鼠脑皮质神经元的损伤机制。方法应用ATP酶测试盒法、荧光分光光度法和高效液相色谱法,分别检测神经细胞线粒体内ATP酶活性、细胞培养上清液中去甲肾上腺素(NA)和多巴胺(DA)含量;电镜下观察MDMA对神经元细胞表面和内部超微结构的组织形态学影响。结果与正常对照组同步比较,培养8d后加入MDMA后继续培养1d,MDMA(50~2000μmol/L)组显示ATP酶活性剂量依赖性降低,但在加入MDMA的细胞培养上清中未检出NA和DA,MDMA(2000μmol/L)组观察到了显著的病理组织形态学改变。结论MDMA对原代培养乳鼠脑皮质神经元有直接损伤作用,该损伤作用非NA和DA介导,与抑制ATP酶活性密切相关。

An experimental study on mechanism of damage to primarily cultured cortical neurons by MDMA in neonate mice

Objective To study the mechanism of damage to cultured cortical neurons by 3,4-methylenedioxymethyl amphetamine (MDMA) in neonate mice. Methods The ATPase activities in the neuronal mitochondria by ATPase assay kit,the content of noradrenaline(NA)by spectrophotofluorimetry and the content of dopamine (DA) in the cellular supernatants by High performance liquid chromatography (HPLC)were measured respectively. The histopathological changes of the cellular surfaces and inner ultrastructures of the neurons were observed under the electronic microscopes. Results As compared with the control group,the dose-dependent reduction of ATPase activities in the MDMA delivered(50～2000 μmol/L)groups,but no detectable NA and DA in the cultured supernatants. Significant histomorphological changes in the MDMA delivered(2000μmol/L)group were observed 1d after administration of MDMA to the neuronal cultures at 8d post-culture. Conclusion MDMA has direct harmful effect on cultured cortical neurons from neonate mice,which is closely associated with its inhibition of ATPase activity,and not mediated by NA and DA.