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Neuroprotection of Interleukin-6 Against NMDA-Induced Apoptosis and Its Signal-Transduction Mechanisms

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Abstract

We have previously shown that interleukin-6 (IL-6)-protected neurons against the suppression of neuronal vitality and overload of intracellular Ca2+ induced by glutamate or N-methyl-d-aspartate (NMDA). Herein we provide further evidence for IL-6 neuroprotection against NMDA-induced apoptosis and explore the signal-transduction mechanisms underlying the anti-apoptotic action of IL-6. Cerebellar granule neurons (CGNs) from postnatal 8-day infant rats were chronically exposed to IL-6 (40 or 120 ng/ml) for 8 days, and stimulated with NMDA (100 μM) for 30 min. To observe the signaling pathways, we employed AG490 (5 or 10 μM), an inhibitor of Janus kinases (JAKs), or LY294002 (5 or 10 μM), an inhibitor of phosphatidylinositol 3-kinase (PI3K), to pretreat the CGNS together with IL-6. The levels of phosphorylation for the downstream effectors of JAKs and PI3K, i.e., phosphorylated STAT3 and Akt, were quantified by Western blot assay. In the cultured CGNs with various drug exposures, the expressions of Bcl-2, Bax, and caspase-3 were measured by real-time PCR and Western blot, and the percentage of apoptotic nuclei was tested by Hoechst 33342 staining. After the CGNs were chronically exposed to IL-6, NMDA stimulation led to an increase in the expression of Bcl-2 mRNA and a decrease in the expression of Bax and caspase-3 mRNAs and proteins when compared with those neurons lacking IL-6 exposure. IL-6 pretreatment of the neurons without NMDA stimulation concentration-dependently enhanced the expressions of Bcl-2 mRNA and protein while attenuating the expressions of Bax and caspase-3 mRNAs and proteins in comparison with control lacking any treatment. Furthermore, IL-6 prevented the increase in the percentage of apoptotic neurons induced by NMDA. The combined pretreatment of the CGNs with AG490 and IL-6 or with LY294002 and IL-6 reduced these anti-apoptotic effects of IL-6. Neither AG490 nor LY294002 exposure alone altered the expressions of Bcl-2, Bax, and cleaved caspase-3 proteins. IL-6 up-regulated the levels of phosphorylated STAT3 and Akt, and this was blocked by AG490 and LY294002, respectively. These results suggest that IL-6 protects neurons against NMDA-induced apoptosis, and that the IL-6 neuroprotection is jointly mediated by JAK-STAT3 and PI3K-Akt signaling pathways.

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Acknowledgments

This study was supported by Grants 2007-5 for Excellent Sci-Tech Innovation Teams in Universities from the Educational Department of Jiangsu Province of China, H200761 from the Health Department of Jiangsu Province of China, 30870819 and 30870929 from the National Natural Science Foundation of China, and K2008006 and K2008019 from the Nantong Applied Research Program of China.

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  1. Department of Physiology, School of Medicine, Nantong University, 19 Qixiu Road, Nantong, 226001, China

    Zhan Liu, Yi-Hua Qiu, Bing Li, Song-Hua Ma & Yu-Ping Peng

  2. The Key Laboratory of Neuroregeneration of Jiangsu Province, Nantong University, 19 Qixiu Road, Nantong, 226001, China

    Yi-Hua Qiu

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  1. Zhan Liu
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Correspondence to Yi-Hua Qiu or Yu-Ping Peng.

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Liu, Z., Qiu, YH., Li, B. et al. Neuroprotection of Interleukin-6 Against NMDA-Induced Apoptosis and Its Signal-Transduction Mechanisms. Neurotox Res 19, 484–495 (2011). https://doi.org/10.1007/s12640-010-9215-x

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