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A Co-Culture Method to Study Neurite Outgrowth in Response to Dental Pulp Paracrine Signals
Published on: February 14, 2020
Initial Insights into the NET-Associated ceRNA Network in Pulpitis: Transcriptomic and Functional Exploration
Xiaolan Guo1, Kailun Wu2, Longrui Dang2
1Shenzhen Clinical College of Stomatology, School of Stomatology, Southern Medical University, Shenzhen, China; Shenzhen Stomatology Hospital (Pingshan) of Southern Medical University, Shenzhen, China.
Background:
Pulpitis is an oral disease primarily triggered by microbial infections. Current therapeutic strategies lack specific agents targeting the underlying pathogenic mechanisms. Non-coding RNAs (ncRNAs) and their competitive endogenous RNA (ceRNA) networks have emerged as critical regulators of diverse biological processes, offering novel insights into the pathogenesis of pulpitis and the identification of potential therapeutic targets.
Methods:
RNA sequencing was performed on pulp tissues from healthy individuals and pulpitis patients. Bioinformatics tools were employed to analyse sequencing data, identify differentially expressed messenger RNAs (mRNAs), and construct protein-protein interaction (PPI) networks to pinpoint hub genes. A murine pulpitis model was established to investigate the effects of neutrophil extracellular trap (NET) inhibitors on disease progression. A NET-associated ceRNA regulatory network was systematically constructed based on ceRNA theory. In vitro experiments validated the expression patterns of key ncRNAs and mRNAs in pulpitis.
Results:
Differentially expressed mRNAs, long non-coding RNAs (lncRNAs), and Circular RNAs (circRNAs) were identified in pulpitis. Bioinformatics analysis revealed significant activation of NET-related pathways, with sialic acid-binding immunoglobulin-like lectin-9 (SIGLEC9), complement C3 (C3), H2A clustered histone 14 (H2AC14), and H4 clustered histone 11 (H4C11) identified as core regulatory genes. In vivo experiments demonstrated that NET inhibition alleviated dental pulpitis and necrosis. Furthermore, a novel NET-associated ceRNA regulatory network was established, revealing four critical regulatory axes: LINC00466 / hsa-miR-27a-3p / SIGLEC9, hsa_circDNAH11_003 / hsa-miR-877-5p / C3, hsa_circSLC15A4_001 / hsa-miR-30d-3p / H2AC14, and hsa_circGABBR2_009 / hsa-miR-193b-3p / H4C11. These ncRNAs exhibited significant upregulation in inflamed pulp tissues.
Conclusion:
By constructing a comprehensive NET-associated ceRNA network, we elucidate molecular mechanisms underlying dental pulpitis, providing novel evidence for understanding the disease progression. These findings establish a theoretical foundation for developing targeted therapeutic strategies against pulpitis.
