牙髓炎中与中性粒细胞胞外陷阱相关的竞争性内源RNA网络的初步见解：转录组学和功能探索

Initial Insights into the NET-Associated ceRNA Network in Pulpitis: Transcriptomic and Functional Exploration.

作者信息

Guo Xiaolan, Wu Kailun, Dang Longrui, Liu Sitong, Xu Jing, Wang Runting, Xu Junyang, Zhong Yiming, Chen Zhao, Wu Buling

机构信息

Shenzhen Clinical College of Stomatology, School of Stomatology, Southern Medical University, Shenzhen, China; Shenzhen Stomatology Hospital (Pingshan) of Southern Medical University, Shenzhen, China.

Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou, China.

出版信息

Int Dent J. 2025 Sep 2;75(6):100958. doi: 10.1016/j.identj.2025.100958.

DOI:10.1016/j.identj.2025.100958

PMID:40902508

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12424408/

Abstract

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.

摘要

背景

牙髓炎是一种主要由微生物感染引发的口腔疾病。目前的治疗策略缺乏针对潜在致病机制的特异性药物。非编码RNA（ncRNA）及其竞争性内源性RNA（ceRNA）网络已成为多种生物学过程的关键调节因子，为牙髓炎的发病机制及潜在治疗靶点的识别提供了新的见解。

方法

对健康个体和牙髓炎患者的牙髓组织进行RNA测序。利用生物信息学工具分析测序数据，鉴定差异表达的信使RNA（mRNA），并构建蛋白质-蛋白质相互作用（PPI）网络以确定枢纽基因。建立小鼠牙髓炎模型以研究中性粒细胞胞外陷阱（NET）抑制剂对疾病进展的影响。基于ceRNA理论系统构建NET相关的ceRNA调控网络。体外实验验证了牙髓炎中关键ncRNA和mRNA的表达模式。

结果

在牙髓炎中鉴定出差异表达的mRNA、长链非编码RNA（lncRNA）和环状RNA（circRNA）。生物信息学分析显示NET相关通路显著激活，唾液酸结合免疫球蛋白样凝集素9（SIGLEC9）、补体C3（C3）、H2A簇组蛋白14（H2AC14）和H4簇组蛋白11（H4C11）被确定为核心调控基因。体内实验表明NET抑制可减轻牙髓炎和牙髓坏死。此外，建立了一个新的NET相关ceRNA调控网络，揭示了四个关键调控轴：LINC00466 / hsa-miR-27a-3p / SIGLEC9、hsa_circDNAH11_003 / hsa-miR-877-5p / C3、hsa_circSLC15A4_001 / hsa-miR-30d-3p / H2AC14和hsa_circGABBR2_009 / hsa-miR-193b-3p / H4C11。这些ncRNA在炎症牙髓组织中显著上调。