Pauwels P J, Van Assouw H P, Peeters L, Moeremans M, Leysen J E
Department of Biochemical Pharmacology, Janssen Research Foundation, Beerse, Belgium.
Synapse. 1992 Dec;12(4):271-80. doi: 10.1002/syn.890120404.
The neuroprotective properties of the cognitive enhancer sabeluzole were investigated in rat brain neuronal cultures derived from the hippocampal formation of 17-day-old rat embryos. Measurement of the neuronal cytoskeletal microtubule-associated protein, MAP2, was used to assess survival of neurons after exposure of neuronal cultures to glutamate. MAP2 was quantified in neuronal cell homogenates by means of an enzyme-linked immunosorbent assay (ELISA) using a mouse monoclonal MAP2 antibody, peroxidase-labeled goat anti-mouse Ig antiserum, and 2,2'-azido-di-[3-ethylbenz-thiazoline] sulphonate (ABTS) as substrate. Exposure of 7-day-old neuronal cultures to 1 mM glutamate for 16 hours led to a three-fold increase in released lactate dehydrogenase (LDH) and a 40% decrease in cellular MAP2 content. Acute treatment of neuronal cultures with 10 microM sabeluzole yielded a 40% drop in released LDH induced by glutamate. Cultures treated chronically with 0.1 microM sabeluzole on days 1 and 4 in culture showed, after 1 week in culture, a MAP2 content and total LDH activity that was not different from control cultures. A 16-hour exposure to 1 mM glutamate did not induce LDH release or changes in MAP2 levels in sabeluzole-treated cultures. A single treatment with 0.1 microM sabeluzole between day 1 to 5 induced a 70-80% drop in glutamate-induced released LDH in 7-day-old neuronal cultures. Full and partial neuronal protection after chronic sabeluzole treatment at 0.1 microM was also observed for neurotoxicity induced by 5 mM N-methyl-D-aspartate (NMDA) and 1 mM kainic acid or 30 microM veratridine, respectively. Within a series of compounds such as Ca++ and Na+ channel antagonists, glutamate receptor antagonists and various neurotransmitter receptor antagonists, sabeluzole, chronically given, were the most potent for inhibition of released LDH induced by 1 mM glutamate (IC50-value: 34 +/- 13 nM). In conclusion, chronic sabeluzole treatment protects cultured rat brain neurons from excitotoxic aggression.
在源自17日龄大鼠胚胎海马结构的大鼠脑神经元培养物中研究了认知增强剂沙贝鲁唑的神经保护特性。通过测量神经元细胞骨架微管相关蛋白MAP2，来评估神经元培养物暴露于谷氨酸后神经元的存活情况。利用小鼠单克隆MAP2抗体、过氧化物酶标记的山羊抗小鼠Ig抗血清以及2,2'-叠氮基二-[3-乙基苯并噻唑啉]磺酸盐（ABTS）作为底物，通过酶联免疫吸附测定（ELISA）对神经元细胞匀浆中的MAP2进行定量。将7日龄的神经元培养物暴露于1 mM谷氨酸中16小时，导致释放的乳酸脱氢酶（LDH）增加了三倍，细胞MAP2含量降低了40%。用10 microM沙贝鲁唑对神经元培养物进行急性处理，可使谷氨酸诱导释放的LDH下降40%。在培养的第1天和第4天用0.1 microM沙贝鲁唑长期处理的培养物，在培养1周后，其MAP2含量和总LDH活性与对照培养物无差异。暴露于1 mM谷氨酸16小时，在经沙贝鲁唑处理的培养物中未诱导LDH释放或MAP2水平变化。在第1天至第5天之间用0.1 microM沙贝鲁唑单次处理，可使7日龄神经元培养物中谷氨酸诱导释放的LDH下降70 - 80%。对于分别由5 mM N-甲基-D-天冬氨酸（NMDA）、1 mM海人酸或30 microM藜芦碱诱导的神经毒性，在0.1 microM长期沙贝鲁唑处理后也观察到了完全和部分的神经元保护作用。在一系列化合物中，如Ca++和Na+通道拮抗剂、谷氨酸受体拮抗剂以及各种神经递质受体拮抗剂，长期给予沙贝鲁唑对抑制1 mM谷氨酸诱导释放的LDH最为有效（IC50值：34±13 nM）。总之，长期沙贝鲁唑处理可保护培养的大鼠脑神经元免受兴奋性毒性攻击。
