Department of Anesthesiology, Wake Forest University School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA.
Eur J Pharmacol. 2012 Feb 29;677(1-3):87-92. doi: 10.1016/j.ejphar.2011.12.015. Epub 2011 Dec 21.
We have recently demonstrated that the glutamate transporter activator riluzole paradoxically enhanced glutamate-induced glutamate release from cultured astrocytes. We further showed that both riluzole and the α(2)δ subunit ligand gabapentin activated descending inhibition in rats by increasing glutamate receptor signaling in the locus coeruleus and hypothesized that these drugs share common mechanisms to enhance glutamate release from astrocytes. In the present study, we examined the effects of riluzole and gabapentin on glutamate uptake and release and glutamate-induced Ca(2+) responses in primary cultures of astrocytes. Riluzole and gabapentin facilitated glutamate-induced glutamate release from astrocytes and significantly increased glutamate uptake, the latter being completely blocked by the non-selective glutamate transporter blocker DL-threo-β-benzyloxyaspartic acid (DL-TBOA). Riluzole and gabapentin also enhanced the glutamate-induced increase in intracellular Ca(2+) concentrations. Some α(2)δ subunit ligands, pregabalin and L-isoleucine, enhanced the glutamate-induced Ca(2+) response, whereas another, 3-exo-aminobicyclo[2.2.1]heptane-2-exo-carboxylic acid (ABHCA), did not. The enhancement of glutamate-induced intracellular Ca(2+) response by riluzole and gabapentin was blocked by the DL-TBOA and an inhibitor of Na(+)/Ca(2+) exchange, 2-[2-[4-(4-nitrobenzyloxy)phenyl]ethyl]isothiurea (KB-R7943). Gabapentin's enhancement of Ca(2+) increase was specific to glutamate stimulation, as it was not mimicked with stimulation by ATP. These results suggest that riluzole and gabapentin enhance Na(+)-glutamate co-transport through glutamate transporters, induce subsequent Ca(2+) influx via the reverse mode of Na(+)/Ca(2+) exchange, and thereby facilitate Ca(2+)-dependent glutamate release by glutamate in astrocytes. The present study also demonstrates a novel target of gabapentinoid action in astrocytes other than α(2)δ subunits in neurons.
我们最近的研究表明,谷氨酸转运体激活剂利鲁唑出人意料地增强了培养星形胶质细胞中谷氨酸诱导的谷氨酸释放。我们进一步表明,利鲁唑和α(2)δ亚基配体加巴喷丁通过增加蓝斑核中的谷氨酸受体信号,激活了下行抑制,并假设这些药物具有增强星形胶质细胞中谷氨酸释放的共同机制。在本研究中,我们检查了利鲁唑和加巴喷丁对原代培养星形胶质细胞中谷氨酸摄取和释放以及谷氨酸诱导的 Ca(2+)反应的影响。利鲁唑和加巴喷丁促进了星形胶质细胞中谷氨酸诱导的谷氨酸释放,并显著增加了谷氨酸摄取,后者被非选择性谷氨酸转运体阻断剂 DL-苏氨酸-β-苄氧基天冬氨酸(DL-TBOA)完全阻断。利鲁唑和加巴喷丁还增强了谷氨酸诱导的细胞内 Ca(2+)浓度增加。一些 α(2)δ 亚基配体,如普瑞巴林和 L-异亮氨酸,增强了谷氨酸诱导的 Ca(2+)反应,而另一种配体 3-外消旋-氨基双环[2.2.1]庚烷-2-外消旋羧酸(ABHCA)则没有。利鲁唑和加巴喷丁增强谷氨酸诱导的细胞内 Ca(2+)反应被 DL-TBOA 和 Na(+)/Ca(2+)交换抑制剂 2-[2-[4-(4-硝基苄氧基)苯基]乙基]异硫脲(KB-R7943)阻断。加巴喷丁对 Ca(2+)增加的增强作用是谷氨酸刺激特异性的,因为它不能模拟 ATP 刺激。这些结果表明,利鲁唑和加巴喷丁通过谷氨酸转运体增强 Na(+)-谷氨酸共转运,诱导随后通过 Na(+)/Ca(2+)交换的反向模式进入 Ca(2+)内流,从而促进星形胶质细胞中谷氨酸依赖的 Ca(2+)依赖的谷氨酸释放。本研究还证明了gabapentinoid 作用在星形胶质细胞中的除神经元的 α(2)δ 亚基以外的新靶点。