Interleukin-10 prevents glutamate-mediated cerebellar granule cell death by blocking caspase-3-like activity
Interleukin-10 prevents glutamate-mediated cerebellar granule cell death by blocking caspase-3-like activity
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DOI：
10.1523/jneurosci.21-09-03104.2001
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发表时间：
2001-05-01
影响因子：
5.3
通讯作者：
Mocchetti, I
中科院分区：
文献类型：
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作者：
Bachis, A;Colangelo, AM;Mocchetti, I
白细胞介素-10（IL-10）已被证明可以减少CNS损伤后的神经元变性。然而，这种细胞因子的神经保护特性的分子机制仍在研究中。谷氨酸会加重创伤引起的继发性损伤。因此，我们检查了IL-10是否阻止谷氨酸介导的细胞死亡。我们在培养中使用大鼠小脑颗粒细胞，因为这些神经元在暴露于毒性浓度的谷氨酸（100-500 μ M）或NMDA（300 μ M）时发生凋亡。用IL-10（1-50 ng/ml）预处理小脑颗粒细胞引起谷氨酸诱导的兴奋性毒性的剂量和时间依赖性降低。最重要的是，IL-10减少凋亡细胞的数量时，添加到一起或谷氨酸后1小时的培养物。利用膜片钳技术和荧光钙离子成像技术，我们研究了IL-10是否通过阻断NMDA通道功能来防止谷氨酸毒性。IL-10不能影响NMDA通道特性，也不能降低NMDA介导的细胞内Ca 2+升高。因此，这种细胞因子似乎通过与NMDA受体功能阻断无关的机制来预防谷氨酸毒性。已经提出各种蛋白酶，如半胱天冬酶-3和转录因子，如核因子κ B（NF-κ B），参与谷氨酸介导的细胞凋亡。因此，我们研究了IL-10是否调节这些凋亡标志物的活性。IL-10阻断了谷氨酸介导的caspase-3诱导以及NF-κ B DNA结合活性，表明IL-10的神经保护特性可能依赖于其阻断促凋亡蛋白活性的能力。
Interleukin-10 (IL-10) has been shown to reduce neuronal degeneration after CNS injury. However, the molecular mechanisms underlying the neuroprotective properties of this cytokine are still under investigation. Glutamate exacerbates secondary injury caused by trauma. Thus, we examined whether IL-10 prevents glutamate-mediated cell death. We used rat cerebellar granule cells in culture because these neurons undergo apoptosis upon exposure to toxic concentrations of glutamate (100-500 muM) or NMDA (300 muM). Pretreatment of cerebellar granule cells with IL-10 (1-50 ng/ml) elicited a dose- and time-dependent reduction of glutamate-induced excitotoxicity. Most importantly, IL-10 reduced the number of apoptotic cells when added to the cultures together or 1 hr after glutamate. Using patch-clamping and fluorescence Ca2+ imaging techniques, we examined whether IL-10 prevents glutamate toxicity by blocking the function of NMDA channel. IL-10 failed to affect NMDA channel properties and to reduce NMDA-mediated rise in intracellular Ca2+. Thus, this cytokine appears to prevent glutamate toxicity by a mechanism unrelated to a blockade of NMDA receptor function. Various proteases, such as caspase-3, and transcription factors, such as nuclear factor kappaB (NF-kappaB), have been proposed to participate in glutamate-mediated apoptosis. Thus, we examined whether IL-10 modulates the activity of these apoptotic markers. IL-10 blocked both the glutamate-mediated induction of caspase-3 as well as NF-kappaB DNA binding activity, suggesting that the neuroprotective properties of IL-10 may rely on its ability to block the activity of proapoptotic proteins.