MicroRNA-181b attenuates lipopolysaccharide-induced inflammatory responses in pulpitis via the PLAU/AKT/NF-κB axis

  • Int Immunopharmacol. 2023 Dec 26:127:111451. doi: 10.1016/j.intimp.2023.111451.
Tiantian Meng  1 ,  Xinpai Liu  2 ,  Jing Zhang  3 ,  Song Li  4 ,  Wei He  5 ,  Wuli Li  6
Affiliations
  • 1. College & Hospital of Stomatology, Anhui Medical University, Key Lab. of Oral Diseases Research of Anhui Province, 69# Mei Shan Road, Hefei 230032, Anhui, China. Electronic address: [email protected].
  • 2. College & Hospital of Stomatology, Anhui Medical University, Key Lab. of Oral Diseases Research of Anhui Province, 69# Mei Shan Road, Hefei 230032, Anhui, China. Electronic address: [email protected].
  • 3. College & Hospital of Stomatology, Anhui Medical University, Key Lab. of Oral Diseases Research of Anhui Province, 69# Mei Shan Road, Hefei 230032, Anhui, China. Electronic address: [email protected].
  • 4. College & Hospital of Stomatology, Anhui Medical University, Key Lab. of Oral Diseases Research of Anhui Province, 69# Mei Shan Road, Hefei 230032, Anhui, China. Electronic address: [email protected].
  • 5. College & Hospital of Stomatology, Anhui Medical University, Key Lab. of Oral Diseases Research of Anhui Province, 69# Mei Shan Road, Hefei 230032, Anhui, China; School of Basic Medical Sciences, Anhui Medical University, 81#Mei Shan Road, Hefei 230032, Anhui, China. Electronic address: [email protected].
  • 6. College & Hospital of Stomatology, Anhui Medical University, Key Lab. of Oral Diseases Research of Anhui Province, 69# Mei Shan Road, Hefei 230032, Anhui, China. Electronic address: [email protected].
PMID: 38154211 DOI: 10.1016/j.intimp.2023.111451
Abstract

Objective: This study aimed to investigate the role and underlying mechanisms of MicroRNA (miRNA)-181b in the inflammatory response in pulpitis.

Methods: Quantitative reverse-transcription polymerase chain reaction (qRT-PCR), fluorescence in situ hybridization (FISH), and immunofluorescence techniques were used to determine the miRNA-181b and urokinase-type plasminogen activator (PLAU) expression levels in inflamed human dental pulp tissues (HDPTs) and lipopolysaccharide (LPS)-stimulated human dental pulp cells (hDPCs). The targets of miRNA-181b were identified and confirmed using a bioinformatics analysis, RNA Sequencing, and dual-luciferase gene reporter assays. The effect of miRNA-181b or PLAU on proinflammatory cytokine expression in hDPCs was examined using qRT-PCR and western blotting. RNA Sequencing was conducted to examine the signaling pathways implicated in miRNA-181b-mediated pulpitis. Western blotting and qRT-PCR were used to determine the miRNA-181b /PLAU/Akt/NF-κB signaling axis in pulpitis. A rat pulpitis model was created to observe the histopathological changes in the dental pulp tissue after the topical application of miRNA-181b agomir.

Results: A significant decrease in miRNA-181b and an increase in PLAU were observed in HDPTs compared to the healthy controls, and these two factors showed a negative correlation. MiRNA-181b directly targeted PLAU. The miRNA-181b inhibitor resulted in a significant upregulation of IL-1β, IL-6 and TNF-α, whereas the knockdown of PLAU reversed this proinflammatory effect. Conversely, PLAU overexpression prevented the anti-inflammatory effects of the miRNA-181b mimics. Mechanistically, miRNA-181b inhibited the Akt/NF-κB pathway by targeting PLAU. In vivo application of the miRNA-181b agomir to inflamed pulp tissue alleviated inflammation.

Conclusion: MiRNA-181b targets PLAU, negatively regulating pro-inflammatory cytokine expression via the Akt/NF-κB signaling pathway.

Keywords
AKT/NF-κB pathway; Inflammation; PLAU; Pulpitis; microRNA-181b.
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