Topiramate protects against glutamate- and kainate-induced neurotoxicity in primary neuronal-astroglial cultures
Topiramate protects against glutamate- and kainate-induced neurotoxicity in primary neuronal-astroglial cultures
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DOI：
10.1016/s0920-1211(03)00039-1
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发表时间：
2003-04-01
影响因子：
2.2
通讯作者：
Hansson, E
中科院分区：
文献类型：
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作者：
Ängehagen, M;Ben-Menachem, E;Hansson, E
抗癫痫药物（AED）托吡酯（TPM）的潜在神经保护作用进行了评估，使用原代神经元星形胶质细胞培养或星形胶质细胞富集培养暴露于兴奋毒性浓度的谷氨酸（Glu）或红藻氨酸的新生大鼠。通过免疫细胞化学和Ca 2+成像评价神经元表达NMDA和AMPA/红藻氨酸盐类型的功能性Gln受体。当将Glu（10 mM）加入到与荧光染料钙黄绿素/AM孵育5小时的9-10天培养物中时，在两种培养类型中均存在显著的细胞损失，但在神经元-星形胶质细胞培养物中更为明显。当TPM（5-10 μ M）与Glu一起包含在培养基中时，在神经元-星形胶质细胞培养物中存活细胞的量显著更高，但在星形胶质细胞富集培养物中不是。培养物的免疫标记显示，当TPM包含在含Glu的培养基中时，MAP阳性神经元细胞的存活率提高。由于TPM对红藻氨酸激活的受体具有已证实的负调节作用，将神经元-星形胶质细胞培养物进一步暴露于兴奋毒性浓度的红藻氨酸（100 μ M）并进行化学分析。显著更多的MAP阳性神经元在含有TPM的培养基中存活，并且显示出与未处理的细胞相似的形态。丙戊酸盐和苯妥英用作参比AED。总之，我们的研究结果表明，TPM对神经元细胞在原代培养，暴露于兴奋性毒性水平的谷氨酸或红藻氨酸的保护作用。(C)2003 Elsevier Science B. V.保留所有权利。
Potential neuroprotective effects of the antiepileptic drug (AED) topiramate (TPM) were evaluated using primary neuronal-astroglial cultures or astroglial-enriched cultures from newborn rats exposed to excitotoxic concentrations of glutamate (Glu) or kainate. Neurons expressed functional Gin receptors of the NMDA and AMPA/kainate types as evaluated by immunocytochemistry and Ca2+ imaging. When Glu (10 mM) was added to 9-10-day cultures incubated with the fluorescent dye calcein/AM for 5 h, there was a marked cell loss in both culture types, but was more pronounced in the neuronal-astroglial cultures. When TPM (5-10 muM) was included in the medium together with Glu, the amount of surviving cells was significantly higher in the neuronal-astroglial cultures, but not in the astroglial-enriched cultures. Immuno-labeling of the cultures revealed an enhanced survival of MAP positive neuronal cells when TPM was included in the Glu containing medium. As TPM has a proven negative modulatory effect on kainate activated receptors, neuronal-astroglial cultures were further exposed to excitotoxic concentrations of kainate (100 muM) and analyzed immunohistochemically. Significantly more MAP positive neurons survived in the TPM containing medium and showed a morphology similar to untreated cells. Valproate and phenytoin were used as reference AEDs. In conclusion, our results demonstrate a protective effect of TPM upon neuronal cells in primary culture, exposed to excitotoxic levels of Glu or kainate. (C) 2003 Elsevier Science B.V. All rights reserved.