Department of Experimental Neuroendocrinology, Institute of Pharmacology, Polish Academy of Sciences, Smetna 12, 31-343, Krakow, Poland,
Neurotox Res. 2009 Jan;15(1):24-37. doi: 10.1007/s12640-009-9002-8. Epub 2009 Feb 4.
One of the serious unwanted effects of the anthracycline anticancer drug doxorubicin (Dox, adriamycin) is its neurotoxicity, which can be evoked by the activation of extracellular (FAS/CD95/Apo-1) pathway of apoptosis in cells. Since memantine, a clinically used N-methyl-D: -aspartic acid (NMDA) receptor antagonist, shows antiapoptotic action in several models of neuronal cell damage, in this study we evaluated the effect of memantine on the cell death induced by Dox in primary neuronal cell cultures. First, we investigated the effect of different concentrations of Dox (0.1-5 microM) on mouse neocortical, hippocampal, striatal, and cerebellar neurons on 7- and 12-day in vitro (DIV). The 7 DIV neuronal cell cultures were more prone to Dox-induced cell death than 12 DIV cultures. The cerebellar neurons were the most resistant to Dox-induced apoptosis in comparison to neuronal cell cultures derived from the forebrain. Memantine (0.1-2 microM) attenuated the Dox-evoked lactate dehydrogenase release in 7 DIV neuronal cell cultures with no significant effect on 12 DIV cultures. The ameliorating effect of memantine on Dox-mediated cell death was also confirmed by an increase in cell viability measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assay. There was no effect of memantine on Dox-induced caspase-8 and -3 activity and Dox-evoked decrease in mitochondrial potential, although attenuation in the number of cells with apoptotic DNA fragmentation was observed. We also showed that the antiapoptotic effect of memantine in our model was NMDA receptor-independent, since two other antagonists of this receptor, MK-801 and AP-5, did not attenuate Dox-induced cell death. Furthermore, memantine did not influence the Dox-evoked increase in cytoplasmic Ca2+ level. The obtained data suggest developmental regulation of both, the Dox-mediated neurotoxicity and efficacy of memantine in alleviating the Dox-induced cell damage in neuronal cell cultures. Moreover, this neuroprotective effect of memantine seems not to be dependent on caspase-3 activity and on the antagonistic action on NMDA receptor.
蒽环类抗癌药物阿霉素（多柔比星，阿霉素）的一种严重的不良反应是其神经毒性，这可以通过细胞外（FAS/CD95/Apo-1）凋亡途径的激活来诱发。由于临床上使用的 N-甲基-D：-天冬氨酸（NMDA）受体拮抗剂美金刚在几种神经元细胞损伤模型中表现出抗凋亡作用，因此在本研究中，我们评估了美金刚对原代神经元细胞培养物中多柔比星诱导的细胞死亡的影响。首先，我们研究了不同浓度的多柔比星（0.1-5μM）对 7 天和 12 天体外培养（DIV）的小鼠大脑皮质、海马、纹状体和小脑神经元的影响。与 12 DIV 培养物相比，7 DIV 神经元细胞培养物更容易受到多柔比星诱导的细胞死亡。与源自前脑的神经元细胞培养物相比，小脑神经元对多柔比星诱导的凋亡最具抵抗力。美金刚（0.1-2μM）可减轻 7 DIV 神经元细胞培养物中多柔比星诱导的乳酸脱氢酶释放，对 12 DIV 培养物无明显影响。通过 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴盐还原测定法测量细胞活力的增加，也证实了美金刚对多柔比星介导的细胞死亡的改善作用。美金刚对多柔比星诱导的 caspase-8 和 -3 活性以及多柔比星诱导的线粒体膜电位降低没有影响，尽管观察到凋亡 DNA 片段化的细胞数量减少。我们还表明，我们模型中美金刚的抗凋亡作用与 NMDA 受体无关，因为该受体的另外两种拮抗剂 MK-801 和 AP-5 不能减轻多柔比星诱导的细胞死亡。此外，美金刚不影响多柔比星诱导的细胞质 Ca2+水平升高。这些数据表明，多柔比星介导的神经毒性和减轻神经元细胞培养物中多柔比星诱导的细胞损伤的美金刚疗效均受发育调控。此外，美金刚的这种神经保护作用似乎不依赖于 caspase-3 活性和 NMDA 受体的拮抗作用。
