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Effects of NLRP6 in Cerebral Ischemia/Reperfusion (I/R) Injury in Rats

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Abstract

The NOD-like receptor protein 6 (NLRP6), an intracytoplasmic pattern recognition receptor in the nucleotide-binding domain, leucine-rich repeat-containing (NLR) innate immune receptor family, influences the inflammation reaction. The role of NLRP6 in cerebral ischemia-reperfusion (I/R) injury in rats is unclear. We explore the function of NLRP6 in cerebral I/R injury. The investigators used a middle cerebral artery occlusion/reperfusion model (MCAO) to imitate ischemic injury. We found the peak expression of NLRP6 is in 48-h post-cerebral I/R injury. The expression of NLRP6 siRNA, as well as the expression of protein and mRNA, was detected by Western blot and qRT-PCR. The degree of IL-1β and IL-18 was assessed by ELISA. After downregulating NLRP6, the expression of IL-1β, IL-18, cleaved Caspase-1, and myeloperoxidase (MPO) were reduced. In HE and Nissl staining, pathological injury of brain tissue after downregulating NLRP6 was improved. NLRP6 siRNA decreased the NLRP6-ASC binding states by CO-IP. NRP6 has a pro-inflammatory effect in cerebral I/R injury, which may provide a new target for the treatment of cerebral I/R injury.

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Abbreviations

NLRP6:

NOD-like receptor protein 6

I/R injury:

Ischemia-reperfusion injury

MCAO:

Middle cerebral artery occlusion/reperfusion model

siRNA:

Small interfering RNA

ASC:

Apoptosis-associated speck-like protein containing a caspase recruitment domain

IL-1β:

Interleukin-1β

IL-18:

Interleukin-18

Caspase-1:

Cysteinyl aspartate–specific proteinase-1

MPO:

Myeloperoxidase

qRT-PCR:

Reverse transcription-polymerase chain reaction

HE staining:

Hematoxylin and eosin staining

ELISA:

Enzyme-linked immunosorbent assay

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Funding

All experiments were supported by The National Natural Science Foundation of China (Nos.81671158 and 81771261), National Youth Foundation of China (No.81701165), the Natural Science Foundation of Chongqing Science and Technology Committee, China (No.cstc2015jcyA10048).

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Authors and Affiliations

  1. Department of Pathophysiology, Institute of Neuroscience, Chongqing Medical University, Road1, Chongqing, 400016, People’s Republic of China

    Changchang Meng, Yueting Wang & Jing Zhao

  2. Department of Pathology, Chongqing Medical University, Chongqing, People’s Republic of China

    Jinyan Zhang & Zhenyu Li

  3. Department of Molecular Medicine, Chongqing Medical University, Chongqing, People’s Republic of China

    Luyu Zhang

Authors
  1. Changchang Meng
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  2. Jinyan Zhang
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  3. Luyu Zhang
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  4. Yueting Wang
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  5. Zhenyu Li
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  6. Jing Zhao
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Contributions

All authors had full access to all the data in the study and all are responsible for the authenticity and accuracy of experimental data. Literature search and experimental design: Changchang Meng, Jingyan Zhang, and Jing Zhao. Access to data: Changchang Meng, Jingyan Zhang, and Jing Zhao. Analysis and acquaintance of data: Changchang Meng, Jingyan Zhang, and Zhenyu Li. Drafting of the manuscript: Changchang Meng, Jingyan Zhang, Yueting Wang, and Zhenyu Li. Revision of the manuscript: Changchang Meng, Jingyan Zhang, and Yueting Wang. Statistical analysis: Changchang Meng, Jingyan Zhang, Yueting Wang, and Luyu Zhang. Sources of funds: Jing Zhao. Technical and material support: Jing Zhao and Luyu Zhang. Experimental supervision: Jing Zhao.

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Correspondence to Jing Zhao.

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Meng, C., Zhang, J., Zhang, L. et al. Effects of NLRP6 in Cerebral Ischemia/Reperfusion (I/R) Injury in Rats. J Mol Neurosci 69, 411–418 (2019). https://doi.org/10.1007/s12031-019-01370-4

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  • DOI: https://doi.org/10.1007/s12031-019-01370-4

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