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Levetiracetam Prevents Perforin Mediated Neuronal Injury Induced by Acute Cerebral Ischemia Reperfusion

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Abstract

The purpose of this study is to explore the neuroprotection mechanism of levetiracetam (LEV) with acute focal cerebral ischemia-reperfusion (I/P) mouse. The cerebral artery I/P animal model was prepared with a middle artery cerebral occlusion method. For drug intervention, mice were intraperitoneally injected with LEV with a dose of either 15 or 150 mg/kg. Neuronal injury was evaluated by measuring the infarct area, apoptosis ratio, and observation of blood–brain barrier ultrastructure with transmission electron microscope. CD8+ antibody and perforin antibody were used to make cross-reference screen through flow cytometry to determine a perforin-positive rate in CD8+ T lymphocytes (PFN + %). Injection of LEV can reduce infarct area, apoptosis ratio, and blood–brain barrier damage 24 h later after acute I/P in WT mice. In vitro, perforin can lower hippocampal neuron viability. In vivo, removing perforin can relieve neuronal injury. High dose injection of LEV (150 mg/kg) can inhibit perforin secreting from CD8+T lymphocytes. In addition, LEV can still protect neurons with perforin knockout mice. Therefore, our results suggested that LEV may contribute to neuron protection after cerebral ischemia reperfusion. The possible mechanism may be related with perforin release. However, we cannot roll out other mechanisms.

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Acknowledgments

This work was supported by the National Nature Science Foundation of China (81271289), Shanghai Science and Technology Commission (No. 11JC1410700), the Fundamental Research Funds for the Central Universities (No. 1507-219-022) and Academic leader Training Program of Pudong Health Bureau of Shanghai, China (grant number PWRd2012-06)

Author Contributions

Conceived and designed the experiments: GL, XW, YZ and YL. Performed the experiments: GL, YZ, YL, LZ, HB, XHX, JJH and XW. Analyzed the data: XW and YZ. Contributed reagents/materials/analysis tools: YZ, YL, LZ, HB, XHX and JJH. Wrote the paper: XW, YZ. Contributed to study conception and design: GL, XW, YZ and YL. Critically revised the manuscript: GL and XW.

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    1. Department of Neurology, East Hospital, Tongji University School of Medicine, 150Jimo Road, Shanghai, 200120, China

      Yue Zhang, Ying Li, Lian Zuo, Huan Bao, Xiahong Xu, Junjie Hao & Gang Li

    2. Department of Neurology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Xuhui District, 200032, Shanghai, China

      Xin Wang

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    1. Yue Zhang
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    Yue Zhang and Ying Li contributed equally to this work.

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    Zhang, Y., Li, Y., Zuo, L. et al. Levetiracetam Prevents Perforin Mediated Neuronal Injury Induced by Acute Cerebral Ischemia Reperfusion. Mol Neurobiol 53, 5480–5491 (2016). https://doi.org/10.1007/s12035-015-9467-9

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