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Long Non-coding RNA ANRIL Downregulation Alleviates Neuroinflammation in an Ischemia Stroke Model via Modulation of the miR-671-5p/NF-κB Pathway

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Abstract

The aim of this study was to investigate the role and underlying mechanism of the long non-coding RNA ANRIL (antisense noncoding RNA in the INK4 locus, ANRIL) in ischemia stroke (IS) injury. Downregulation of ANRIL by right intracerebroventricular injected si-ANRIL in middle cerebral artery occlusion-reperfusion (MCAO/R) C57/BL6 mice and by transferring si-ANRIL in oxygen glucose deprivation/reperfusion (OGD/R) HT22 cells. The results showed that ANRIL levels increased in IS model, downregulation of ANRIL reduced infract area, neurological deficit scores and injured cells, and prolong fall latency time in MCAO/R mice, improved cell viability and reduced cell cytotoxicity in OGD/R cells. Fluorescence in Situ Hybridization detected that there were both ANRIL and miR-671-5p in neurons; miranda v3.3a and dual luciferase reporter assay demonstrated that miR-671-5p was one of direct target of ANRIL; and our previously published research demonstrated that NF-κB was one of direct target of miR-671-5p. Downregulation of ANRIL alleviated neuroinflammation and reduced p-NF-κB, NF-κB, pro-inflammatory cytokines (IL-1β, IL-6, TNF-a), and iNOS, which diminished by miR-671-5p antagomir both in in vivo and in vitro IS models. Downregulation of ANRIL alleviated disruption of blood brain barrier, and protected against tight junction (ZO-1, occludin and claudin 5) disorder in MCAO/R mice. This work clarified that downregulation of ANRIL reduced neuroinflammation by negatively regulating miR-671-5p to inhibit NF-κB in IS models, which provided a theoretical foundation for the protective effect of downregulating ANRIL for IS patients.

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Data Availability

All data generated or analyzed are available from the corresponding author by emailing 100798@cqmu.edu.cn.

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Acknowledgements

We thank all editors and reviewers for their patience and their valuable comments on this manuscript. We thank LetPub (www.letpub.com) for its linguistic assistance during the preparation of this manuscript.

Funding

This work was funded by Research Innovation of Graduate Students in Chongqing (CYB19165), Natural Science Foundation of Chongqing (cstc2019jcyjmsxmX0630), Natural Science Foundation of Chongqing (cstc2020jcyj-msxmX0325), Chinese Medicine Science and Technology project of Chongqing Municipal Health committee, Grand No. 2021ZY3794 and General Topics of Basic Research and Frontier Exploration of Yuzhong district, Chongqing (20210121).

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  1. The Key Laboratory of Biochemistry and Molecular Pharmacology, College of Pharmacology, Chongqing Medical University, Chongqing, 400016, China

    Ling Deng, Sha Chen, Tianrui Zuo, Qingwen Hu, Yu Wu, Xiaomei Fan & Zhi Dong

  2. Department of Neurology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, People’s Republic of China

    Jin Jiang

  3. Department of Biological Immunotherapy, Chongqing University Cancer Hospital, Shapingba District, Chongqing, 400030, People’s Republic of China

    Xing Lin

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  1. Ling Deng
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Deng, L., Jiang, J., Chen, S. et al. Long Non-coding RNA ANRIL Downregulation Alleviates Neuroinflammation in an Ischemia Stroke Model via Modulation of the miR-671-5p/NF-κB Pathway. Neurochem Res 47, 2002–2015 (2022). https://doi.org/10.1007/s11064-022-03585-1

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