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Ifenprodil Improves Long-Term Neurologic Deficits Through Antagonizing Glutamate-Induced Excitotoxicity After Experimental Subarachnoid Hemorrhage

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Abstract

Excessive glutamate leading to excitotoxicity worsens brain damage after SAH and contributes to long-term neurological deficits. The drug ifenprodil is a non-competitive antagonist of GluN1-GluN2B N-methyl-d-aspartate (NMDA) receptor, which mediates excitotoxic damage in vitro and in vivo. Here, we show that cerebrospinal fluid (CSF) glutamate level within 48 h was significantly elevated in aSAH patients who later developed poor outcome. In rat SAH model, ifenprodil can improve long-term sensorimotor and spatial learning deficits. Ifenprodil attenuates experimental SAH-induced neuronal death of basal cortex and hippocampal CA1 area, cellular and mitochondrial Ca2+ overload of basal cortex, blood-brain barrier (BBB) damage, and cerebral edema of early brain injury. Using in vitro models, ifenprodil declines the high-concentration glutamate-mediated intracellular Ca2+ increase and cell apoptosis in primary cortical neurons, reduces the high-concentration glutamate-elevated endothelial permeability in human brain microvascular endothelial cell (HBMEC). Altogether, our results suggest ifenprodil improves long-term neurologic deficits through antagonizing glutamate-induced excitotoxicity.

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Funding

This study was financially supported by the National Natural Science Foundation of China (82071303, 81870938), Natural Science Foundation of Shandong province (ZR2019ZD32) and Inner Mongolia province (2020MS08167), Youth Innovation Team of Shandong Universities (2019KJK001), Taishan Scholar Project, Science Technology Development Program (2019Z3009-9) of Baotou city, and Academic Promotion Program of Shandong First Medical University (2019QL016, 2019PT007).

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Authors and Affiliations

  1. Department of Orthopedics, Shandong Provincial Hospital Affiliated to Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan, 250021, Shandong, China

    Jing-yi Sun

  2. Department of Neurology, Second Affiliated Hospital; Key Laboratory of Cerebral Microcirculation, Shandong First Medical University & Shandong Academy of Medical Sciences, Yingsheng East Road No.2, Taian, 271016, China

    Jing-yi Sun, Hong-bin Wang, Ya-jun Hou, Qiong-jie Mi, Ming-feng Yang, Hui Yuan, Bao-liang Sun & Zong-yong Zhang

  3. Department of Neurology, Baotou Central Hospital, Baotou, 014040, Inner Mongolia, People’s Republic of China

    Shi-jun Zhao

  4. Postdoctoral Workstation, Taian City Central Hospital, Taian, 271000, Shandong, China

    Qing-bin Ni

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  1. Jing-yi Sun
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Contributions

BS and ZZ designed experiments. JS, SZ, HW, YH, QM, MY, HY, and QN performed experiments. ZZ and BS analyzed the results. ZZ wrote the manuscript with contribution from BS. All authors read and approved the final manuscript.

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Correspondence to Bao-liang Sun or Zong-yong Zhang.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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ESM 1

Ifenprodil (Ifen) reduced high-concentration glutamate (glu)-induced cell damage in rat primary cortical neurons. a Concentration-response curve showing Ca2+ release induced by glutamate alone or together with ifenprodil (100 μM) in cortical neurons. b Effect of ifenprodil (100 μM) on glutamate (50 or 100 μM)-induced Ca2+ release in cortical neurons. c, d Neurons were treated with glutamate (0, 10, 30, 50, 100 or 300 μM) or co-incubation with ifenprodil (100 μM) for 24 h and subsequent determination of the cell viability by CCK8 assay. e Neurons were treated with the glutamate (50 or 100 μM) or co-incubation with ifenprodil (100 μM) for 24 h, then apoptotic neurons were detected by TUNEL assay. Representative photos of six groups were captured from fluorescence microscope (200 ×). Data are mean ± SD. *P < 0.05 vs control, #P < 0.05 vs glu; ns denotes no significant; n = 6. (PNG 6380 kb)

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Sun, Jy., Zhao, Sj., Wang, Hb. et al. Ifenprodil Improves Long-Term Neurologic Deficits Through Antagonizing Glutamate-Induced Excitotoxicity After Experimental Subarachnoid Hemorrhage. Transl. Stroke Res. 12, 1067–1080 (2021). https://doi.org/10.1007/s12975-021-00906-4

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