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NFAT5 protects astrocytes against oxygen–glucose–serum deprivation/restoration damage via the SIRT1/Nrf2 pathway

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Abstract

Nuclear factor of activated T cells (NFAT) is a multifunctional cytokine family. NFAT5 was recently reported to be involved in many neuronal functions, but its specific function remains unclear. In this study, our aim is to investigate whether NFAT5 overexpression can protect astrocytes against oxygen–glucose–serum deprivation/restoration (OGSD/R) damage. In vivo, rats were subjected to ischemia–reperfusion injury, resulting in increased water content, infarct volume, and expression of NFAT5 protein in rat spinal cord. After primary culture for spinal cord astrocytes, the in vitro OGSD/R model was established. The results of the CCK8 assay and flow cytometry showed that, in the OGSD/R group, astrocyte cell viability was downregulated, but astrocyte apoptosis increased. Caspase 3 activity increased as well. Levels of NFAT5, as detected by real-time quantitative PCR and western blot, decreased under OGSD/R, as did SIRT1. Commercial kits for activity assays were used to show that OGSD/R inhibited SIRT1 activation but accelerated SOD activation after OGSD/R. Next, pcDNA-NFAT5 or NFAT5 siRNA was transfected into astrocytes. Overexpression of NFAT5 not only promoted the survival of the astrocytes and SIRT1 activation under OGSD/R but also inhibited cell apoptosis and SOD activation. Moreover, overexpression of NFAT5 apparently diminished histone acetylation and promoted the nuclear transport of Nrf2. Our results show that NFAT5 protects spinal astrocytes in a manner that depends on activation of the SIRT1/Nrf2 pathway. These findings present a novel potential molecular mechanism for NFAT5 therapy in the context of spinal cord injury.

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Abbreviations

NFAT:

Nuclear factor of activated T cells

OGSD/R:

Oxygen–glucose–serum deprivation/restoration

SCI:

Spinal cord injury

I/R:

Ischemia–reperfusion

SIRT1:

Sirtuin 1

HSP70:

Heat shock protein 70

HO-1:

Hemeoxygenase1

Nrf2:

Nuclear factor-E2 related factor 2

Keap1:

Kelch-like ECH-associated protein 1

TTC:

2,3,5-triphenyltetrazolium chloride

siRNA:

Small interfering RNA

GFAP:

Glial fibrillary acid protein

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    Authors and Affiliations

    1. Department of Neurological Surgery, Chengdu Military General Hospital, Chengdu, 610083, China

      Xun Xia, Yun-Ming Li, Li-Bin Yang, Ke-Xia Fan, Hui Zheng, Hai-Dong Huang, Yong-Qin Kuang & Yuan Ma

    2. Department of Orthopaedics, Chengdu Military General Hospital, Chengdu, 610083, China

      Bo Qu

    3. Department of Neurological Surgery, The 306th Hospital of PLA, No. 9 Anxiangbeili, Chaoyang District, Beijing, 100101, China

      Jian-Wen Gu

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    1. Xun Xia
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    2. Bo Qu
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    Correspondence to Jian-Wen Gu, Yong-Qin Kuang or Yuan Ma.

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    Xun Xia, Bo Qu and Yun-Ming Li contributed equally to this work.

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    Xia, X., Qu, B., Li, YM. et al. NFAT5 protects astrocytes against oxygen–glucose–serum deprivation/restoration damage via the SIRT1/Nrf2 pathway. J Mol Neurosci 61, 96–104 (2017). https://doi.org/10.1007/s12031-016-0849-x

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