Research Paper Volume 13, Issue 19 pp 23004—23019
lncRNA MEG3 aggravated neuropathic pain and astrocyte overaction through mediating miR-130a-5p/CXCL12/CXCR4 axis
- 1 Department of Anesthesiology, Qianjiang Hospital Affiliated to Renmin Hospital of Wuhan University, Qianjiang 433100, Hubei, China
- 2 Department of Anesthesiology, The First People's Hospital of Jingzhou, Jingzhou 434000, Hubei, China
- 3 Department of Pain, The First People's Hospital of Jingzhou, Jingzhou 434000, Hubei, China
Received: June 15, 2021 Accepted: September 20, 2021 Published: October 5, 2021https://doi.org/10.18632/aging.203592
How to Cite
Copyright: © 2021 Dong et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Objective: Long non-coding RNAs (lncRNAs) exert a critical function in mediating neuropathic pain (NP). MEG3, a novel lncRNA, contributes to astrocyte activation and inflammation. However, its role in NP remains unclear.
Methods: The chronic constriction injury (CCI) method was employed to construct an NP rat model. Astrocyte activation was induced by lipopolysaccharide (LPS). The profiles of MEG3, microRNA (miR)-130a-5p, CXC motif chemokine receptor 12 (CXCL12)/CXC motif chemokine receptor 4 (CXCR4), and the Rac1/NF-κB pathway in CCI rats’ spinal cord tissues and astrocytes were monitored by reverse transcription-quantitative PCR (RT-qPCR) and western blot (WB). Pain scores of CCI rats were assessed. Enzyme-linked immunosorbent assay (ELISA) was adopted to monitor neuroinflammation alteration. The glial fibrillary acidic protein (GFAP)-labeled astrocytes were tested by immunohistochemistry (IHC). Bioinformatics, dual-luciferase reporter assay and RNA immunoprecipitation (RIP) were utilized to verify the molecular mechanism between MEG3 and miR-130a-3p.
Results: MEG3, CXCL12 and CXCR4 were overexpressed and miR-130a-5p was knocked down in CCI rats and LPS-induced astrocytes. Up-regulating MEG3 aggravated NP, enhanced inflammatory cytokines interleukin-1β (IL-1β), tumor necrosis factor (TNF)-α, and interleukin-6 (IL-6) expression and release in CCI rats and LPS-induced astrocytes. Up-regulating miR-130-5p repressed LPS-induced inflammation in astrocytes. AS verified by the dual-luciferase reporter assay and RIP assay, MEG3 sponged miR-130a-5p as a competitive endogenous RNA (ceRNA). What’s more, miR-130a-5p up-regulation weakened the MEG3-induced proinflammatory effects on LPS-induced astrocytes.
Conclusions: MEG3 aggravates NP and astrocyte activation via the miR-130a-5p/CXCL12/CXCR4 axis, which is a potential therapeutic target for NP.
NP: Neuropathic pain; CCI: chronic constriction injury; LPS: lipopolysaccharide; miR: microRNA; RT-qPCR: reverse transcription-quantitative PCR; WB: western blot; ELISA: enzyme-linked immunosorbent assay; IHC: immunohistochemistry; RIP: immunoprecipitation; IL-6: interleukin-6; lncRNAs: non-coding RNAs; SNL: spinal nerve ligation; MEG3: Maternal expression gene 3; MCAO: middle cerebral artery occlusion; OGD: oxygen-glucose deprivation; PWT: paw withdrawal threshold; PWL: paw withdraw latency; RT: room temperature; TUNEL: TdT-mediated dUTP nick end labeling; AI: apoptosis index; FBS: fetal bovine serum; ATCC: American Type Culture Collection; NHAs: Normal human astrocytes; qRT-PCR: quantitative polymerase chain reaction; ELISA: Enzyme-linked immunosorbent assay; TBI: traumatic brain injury; AD: Alzheimer's disease; SCI: spinal cord injury.