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MiR-181a-5p Alleviates the Inflammatory Response of PC12 Cells by Inhibiting High-Mobility Group Box-1 Protein
Zhiwu Wu1, Zhixiong Zhang1, Zhihua Wang1
1Department of Neurosurgery＆Jiangxi Key Laboratory of Neurosurgery, The First Affiliated Hospital Of Nanchang University, Nanchang, Jiangxi, PR China.
Objective:
Neuroinflammation triggers sequelae after spinal cord injury (SCI). Inhibition of inflammation promotes recovery after SCI. MicroRNAs regulate many pathophysiological processes, including inflammation. Any role for miR-181a-5p in the inflammatory response after SCI remains unclear. Thus, we evaluated the effects of miR-181a-5p on inflammation in PC12 cells and the underlying mechanism in play.
Methods:
Quantitative reverse transcription-polymerase chain reaction was used to measure the levels of miR-181a-5p and high-mobility group box-1 protein (HMGB1) in SCI tissues. Cell-counting kit-8 assays were used to assess the viability of PC12 cells treated with lipopolysaccharide (LPS). Plasmids encoding MiR-181a-5p mimics, an miR-181a-5p inhibitor, or/and the HMGB1 were transfected into PC12 cells. Quantitative reverse transcription-polymerase chain reaction or/and Western blotting were performed to assess the expression of miR-181a-5p, HMGB1, and inflammatory factors in vitro.
Results:
MiR-181a-5p expression decreased and HMGB1 expression increased in SCI tissues and LPS-induced PC12 cells. Upregulation of miR-181a-5p (via transfection) inhibited inflammation of, and HMGB1 expression by, LPS-induced PC12 cells. HMGB1 overexpression reversed the anti-inflammatory effects of miR-181a-5p. Dual-luciferase assays confirmed that HMGB1 was a direct target of miR-181a-5p.
Conclusions:
miR-181a-5p attenuated the inflammatory response of LPS-induced PC12 cells by directly inhibiting HMGB1; thus, miR-181a-5p may serve as a therapeutic target in SCI.
Insights
MicroRNA-181a-5p (miR-181a-5p) reduces inflammation after spinal cord injury (SCI) by targeting high-mobility group box-1 protein (HMGB1). This finding suggests miR-181a-5p as a potential therapeutic target for SCI recovery.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Neuroinflammation is a key factor in spinal cord injury (SCI) pathology.
- Inhibiting inflammation is crucial for promoting recovery after SCI.
- MicroRNAs (miRNAs) are regulators of inflammatory processes, but the role of miR-181a-5p in SCI-induced inflammation is not well understood.
Purpose of the Study:
- To investigate the role of miR-181a-5p in the inflammatory response following spinal cord injury (SCI).
- To elucidate the underlying molecular mechanism by which miR-181a-5p influences inflammation in PC12 cells.
Main Methods:
- Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting were used to measure miR-181a-5p and high-mobility group box-1 protein (HMGB1) levels.
- PC12 cells were treated with lipopolysaccharide (LPS) and transfected with miR-181a-5p mimics, inhibitors, or HMGB1 constructs.
- Cell viability was assessed using Cell-Counting Kit-8 assays, and inflammatory factor expression was analyzed.
Main Results:
- miR-181a-5p expression was decreased, while HMGB1 expression was increased in SCI tissues and LPS-treated PC12 cells.
- Upregulation of miR-181a-5p significantly inhibited inflammation and HMGB1 expression in LPS-induced PC12 cells.
- Overexpression of HMGB1 reversed the anti-inflammatory effects of miR-181a-5p, and dual-luciferase assays confirmed HMGB1 as a direct target of miR-181a-5p.
Conclusions:
- miR-181a-5p attenuates the inflammatory response in LPS-induced PC12 cells by directly inhibiting HMGB1.
- miR-181a-5p demonstrates potential as a therapeutic target for managing inflammation in spinal cord injury.
