Long noncoding RNA ANRIL knockdown attenuates neuroinflammation following ischemic stroke via suppressing the expression of NF-κB in vitro and in vivo

ABSTRACT

Objective

Increasing evidence suggests that long-noncoding RNAs can exert neuroprotective effects in cerebral ischemia-reperfusion injury. Levels of the long noncoding RNA ANRIL (ANRIL) are reportedly altered in ischemic stroke (IS) patients, but its role in IS requires further clarification. This study was designed to explore the mechanistic function of ANRIL in IS.

Methods

In vitro, HT22 cells was treated with an oxygen-glucose deprivation/reperfusion (OGD/R). In vivo, brain ischemia/reperfusion was induced by 60-minute transient middle cerebral artery occlusion/reperfusion (MCAO/R) IS model in C57/BL6 mice. Additionally, cells were transfected with si-ANRIL, pcDNA3.1-ANRIL, pcDNA3.1-NF-κB, or appropriate negative controls, and si-ANRIL and pcDNA3.1-NF-κB were administered into the lateral ventricles in MCAO/R model mice. Cell viability and apoptosis were detected via MTT and flow cytometry assays. mRNA and protein expression of NF-κB were detected via qRT-PCR and Western blotting. IL-1β, IL-6, TNF-a, and iNOS levels were detected via ELISA. In addition, infarcted area and neuronal injury were evaluated via TTC, Nissl, and immunofluorescent staining.

Results

We found that ANRIL knockdown increased cell viability and reduced apoptosis in vitro. Additionally, we found that ANRIL knockdown decreased p-P65, P65, IL-1β, IL-6, TNF-a, and iNOS levels, whereas these effects were reversed by NF-κB overexpression both in vitro and in vivo.

Conclusion

our results suggest that ANRIL knockdown attenuates neuroinflammation by suppressing the expression of NF-κB both in vitro and vivo model of IS, sugguesting that ANRIL might be a potentially viable therapeutictarget to diminish neuroinflammation in IS patients.

GRAPHICAL ABSTRACT

KEYWORDS:

• Long nocoding RNA ANRIL
• neuroinflammation
• NF-κB
• MCAO/R
• cerebral ischemia/reperfusion

Acknowledgments

This study was supported by General Topics of Basic Research and Frontier exploration in chongqing (stc2018jcyjAX0378) and Research Innovation of Graduate Students in Chongqing (CYB19165) .

We thank all editors and reviewers for their help and patience.

Compliance with Ethical Standards

Animals were treated in accordance with animal ethics standards throughout the animal experiment.

Consent for Publication

All authors agree to publish.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by the General Topics of Basic Research and Frontier exploration in chongqing [stc2018jcyjAX0378]; Research Innovation of Graduate Students in Chongqing [CYB19165].

Notes on contributors

Ling Deng

Ling Deng is a PhD candicate of neuropharmacoloy from College of Pharmacology, Chongqing Medical university. She participated in conceptualization, methodology, validation, software, formal analysis, investigation, data curation, funding acquisition, writing - original draft, project administration, supervision and visualization. All authors read and approved the final manuscript.

Yi Guo

Yi Guo is a radiologist from Department of Radiology, Chongqing University Central Hospital. She participated in formal analysis, investigation and resources. All authors read and approved the final manuscript.

Jingdong Liu

Jingdong Liu, Xuan Wanga, Hongxia Zhao, Tianrui Zuo and Qingwen Hu are master of neuropharmacoloy from College of Pharmacology, Chongqing Medical university. They participated in methodology, validation, investigation, supervision, software. All authors read and approved the final manuscript.

Sha Chen

Sha Chen is a doctor of neuropharmacoloy from College of Pharmacology, Chongqing Medical university. She is participated in funding acquisition, writing - review & editing. All authors read and approved the final manuscript.

Zhi Dong

Zhi Dong is a doctor of pharmacology from College of pharmacology, Chongqing Medical university. And he is a doctoral supervisor. His field of researches are on neuroscience and neuropharmacology. He participated in conceptualization, funding acquisition, writing - review & editing and resources.