Open AccessLncRNA NEAT1 alleviates ischemic stroke via transcriptional inhibition of NLRP3 mediated by the miR-10b-5p/BCL6 axis
Author(s) -
Zhiwen Zhou,
Xiang Ren,
Wensheng Zhou,
Aiping Li,
Zheng Liu
Publication year - 2022
Publication title -
acta neurobiologiae experimentalis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.542
H-Index - 55
eISSN - 1689-0035
pISSN - 0065-1400
DOI - 10.55782/ane-2022-002
Subject(s) - inflammasome , downregulation and upregulation , western blot , inflammation , chemistry , luciferase , neuroinflammation , bcl6 , microglia , microbiology and biotechnology , cancer research , biology , transfection , immunology , gene , biochemistry , antibody , germinal center , b cell
Cerebral ischemic stroke (CIS) is a significant cause of disability and death. Inflammation usually occurs after CIS and acceleratescellular damage. NLRP3 plays a key role in the formation of CIS‑associated inflammasome. Understanding how NLRP3 is regulatedbears great importance. We hypothesized that lncRNA NEAT1 can downregulate NLRP3 expression by regulating the miR‑10b‑5p/BCL6axis, and thus regulate microglia‑driven inflammation. The expression of NEAT1 was analyzed in CIS patients and an in vitro modelof oxygen and glucose deprivation/re‑oxygenation (OGD/R). We assessed the levels of pro‑inflammatory cytokines IL‑18 and IL‑1βwith ELISA. Interactions between NEAT1/miR‑10b‑5p and miR‑10b‑5p/BCL6 were determined by luciferase assay. The interaction ofBCL6 and NLRP3 was identified by ChIP; RNA, and protein levels were evaluated by qRT‑PCR and western blot, respectively. We foundthat NEAT1 level was decreased in CIS patients and OGD/R treated cells. OGD/R exerted pro‑inflammasome effects by increasing theexpression of inflammasome‑associated proteins and ROS and malondialdehyde (MDA) while inhibiting SOD production. This effectwas partially antagonized by NEAT1. We bioinformatically identified interactions between NEAT1/miR‑10b‑5p, BCL6/miR‑10b‑5p, andNLRP3‑promoter/BCL6, and validated them by luciferase assay, qRT‑PCR, and ChIP. NEAT1 inhibited miR‑10b‑5p and upregulated BCL6by ceRNA mechanism and alleviated OGD/R induced cell damage. We also proved that BCL6 was a repressive transcription factor inthe regulation of NLRP3 expression. Thus, lncRNA NEAT1 inhibited inflammasome activation by NLRP3 in microglia via the NEAT1/miR‑10b‑5p/BCL6/NLRP3 regulatory axis, which alleviated deleterious outcomes of ischemic stroke.