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Impact of Heat Shock Protein A 12B Overexpression on Spinal Astrocyte Survival Against Oxygen-Glucose-Serum Deprivation/Restoration in Primary Cultured Astrocytes

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Abstract

Heat shock protein A 12B (HSPA12B) is a newly discovered member of the heat shock protein 70 family. Preclinical evidence indicates that HSPA12B helps protect the brain from ischemic injury, although its specific function remains unclear. The aim of this study is to investigate whether HSPA12B overexpression can protect astrocytes from oxygen-glucose-serum deprivation/restoration (OGD/R) injury. We analyzed the effects of HSPA12B overexpression on spinal cord ischemia-reperfusion injury and spinal astrocyte survival. After ischemia-reperfusion injury, we found that HSPA12B overexpression decreased spinal cord water content and infarct volume. MTT assay showed that HSPA12B overexpression increased astrocyte survival after OGD/R treatment. Flow cytometry results showed a marked inhibition of OGD/R-induced astrocyte apoptosis. Western blot assay showed that HSPA12B overexpression significantly increased regulatory protein B-cell lymphocyte 2 (Bcl-2) levels, whereas it decreased expression of the Bax protein, which forms a heterodimer with Bcl-2. Measurements of the level of activation of caspase-3 by Caspase-Glo®3/7 Assay kit showed that HSPA12B overexpression markedly inhibited caspase-3 activation. Notably, we demonstrated that the effects of HSPA12B on spinal astrocyte survival depended on activation of the PI3K/Akt signal pathway. These findings indicate that HSPA12B protects against spinal cord ischemia-reperfusion injury and may represent a potential treatment target.

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    Authors and Affiliations

    1. Postgraduate Department, Third Military Medical University, Chongqing, 400038, China

      Xun Xia

    2. Department of Neurological Surgery, Chengdu Military General Hospital, No.270 Rongdu Road, Chengdu, Sichuan Province, 610083, China

      Xun Xia, Yuan Ma, Li-bin Yang, Jing-ming Cheng, Tao Yang, Ke-xia Fan, Yun-ming Li, En-yu Liu, Lin Cheng, Hai-dong Huang & Yong-qin Kuang

    3. Department of Neurological Surgery, The 306th Hospital of PLA, N0.9 Anxiang Beili, Chaoyang District, Beijing, 100101, China

      Jian-wen Gu

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    1. Xun Xia
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    Correspondence to Jian-wen Gu or Yong-qin Kuang.

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    All experimental protocols were carried out by the Institutional Animal Care at Third Military Medical University and were approved by the Ethics Committee for Animal Experimentation of Third Military Medical University, Chongqing, China. Every effort was made in this study to minimize the number of rats used and their suffering during experiments.

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    Xun Xia and Yuan Ma contributed equally to this work.

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    Xia, X., Ma, Y., Yang, Lb. et al. Impact of Heat Shock Protein A 12B Overexpression on Spinal Astrocyte Survival Against Oxygen-Glucose-Serum Deprivation/Restoration in Primary Cultured Astrocytes. J Mol Neurosci 59, 511–520 (2016). https://doi.org/10.1007/s12031-016-0768-x

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