Kainic acid-induced neuronal cell death in cerebellar granule cells is not prevented by caspase inhibitors
作者:Ester Verdaguer、Elvira García-Jordà、Andrés Jiménez、Alessandra Stranges、Francesc X Sureda、Anna M Canudas、Elena Escubedo、Jordi Camarasa、Mercè Pallàs、Antoni Camins
DOI:10.1038/sj.bjp.0704581
日期:2002.3
We examined the role of non‐NMDA receptors in kainic acid (KA)‐induced apoptosis in cultures of rat cerebellar granule cells (CGCs). KA (1 – 500 μM) induced cell death in a concentration‐dependent manner, which was prevented by NBQX and GYKI 52466, non‐NMDA receptor antagonists. Moreover, AMPA blocked KA‐induced excitotoxicity, through desensitization of AMPA receptors.
Similarly, KA raised the intracellular calcium concentration of CGCs, which was inhibited by NBQX and GYKI 52466. Again, AMPA (100 μM) abolished the KA (100 μM)‐induced increase in intracellular calcium concentration.
KA‐induced cell death in CGCs had apoptotic features, which were determined morphologically, by DNA fragmentation, and by expression of the prostate apoptosis response‐4 protein (Par‐4).
KA (500 μM) slightly (18%) increased caspase‐3 activity, which was strongly enhanced by colchicine (1 μM), an apoptotic stimulus. However, neither Z‐VAD.fmk, a pan‐caspase inhibitor, nor the more specific caspase‐3 inhibitor, Ac‐DEVD‐CHO, prevented KA‐induced cell death or apoptosis. In contrast, both drugs inhibited colchicine‐induced apoptosis.
The calpain inhibitor ALLN had no effect on KA or colchicine‐induced neurotoxicity.
Our findings indicate that colchicine‐induced apoptosis in CGCs is mediated by caspase‐3 activation, unlike KA‐induced apoptosis.
British Journal of Pharmacology (2002) 135, 1297–1307; doi:10.1038/sj.bjp.0704581
我们研究了非NMDA受体在Kainic酸(KA)诱导的大鼠小脑颗粒细胞(CGCs)凋亡中的作用。
- KA(1–500 μM)以浓度依赖性方式诱导细胞死亡,这种作用被非NMDA受体拮抗剂NBQX和GYKI 52466所阻断。此外,AMPA通过使AMPA受体脱敏,阻止了KA诱导的兴奋性毒性。
- 类似地,KA升高了CGCs的胞内钙浓度,这种升高被NBQX和GYKI 52466抑制。此外,AMPA(100 μM)消除了KA(100 μM)引起的胞内钙浓度升高。
- KA诱导的CGCs凋亡具有典型的凋亡特征,包括形态学变化、DNA片段化以及前列腺凋亡反应蛋白-4(Par-4)的表达。
- KA(500 μM)轻微(18%)增加了Caspase-3的活性,而这种活性在加入秋水仙素(1 μM,一种凋亡刺激因子)后显著增强。然而,无论是广谱Caspase抑制剂Z-VAD.fmk,还是更特异的Caspase-3抑制剂Ac-DEVD-CHO,都无法阻止KA诱导的细胞死亡或凋亡。相反,这两种药物均抑制了秋水仙素诱导的凋亡。
- 胰蛋白酶抑制剂ALLN对KA或秋水仙素诱导的神经毒性无影响。
- 我们的发现表明,在CGCs中,秋水仙素诱导的凋亡是由Caspase-3活化介导的,而KA诱导的凋亡则不然。
——《British Journal of Pharmacology》(2002)第135卷,1297–1307页; doi:[10.1038/sj.bjp.0704581](https://doi.org/10.1038/sj.bjp.0704581)