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LncRNA MIAT 在牙周炎中的诊断意义及通过 miR-204-5p/DKK1 轴影响牙周膜成纤维细胞的分子机制。

Diagnostic significance of LncRNA MIAT in periodontitis and the molecular mechanisms influencing periodontal ligament fibroblasts via the miR-204-5p/DKK1 axis.

机构信息

Department of stomatology, Dental Well Institute of Temporomandibular Joint Research, Chengdu, China; LESHAN Vocational and Technical College, Leshan, China.

LESHAN Vocational and Technical College, Leshan, China; Department of stomatology, Leshan Weiduo Dental, Leshan, China.

出版信息

Arch Oral Biol. 2024 Dec;168:106066. doi: 10.1016/j.archoralbio.2024.106066. Epub 2024 Aug 4.

Abstract

OBJECTIVE

This study investigated the clinical importance of long noncoding RNA myocardial infarction-associated transcript (MIAT) in periodontitis and its impact on the functional regulation of human periodontal ligament fibroblasts (hPDLFs).

METHODS

Ninety-eight periodontitis patients and 74 healthy controls were enrolled. In vitro cellular models were created using Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) to stimulate hPDLFs. Real-time quantitative polymerase chain reaction was used to measure mRNA levels of MIAT and osteogenic factors. Inflammation factor concentration was assessed using an enzyme-linked immunosorbent assay. Cell viability and apoptosis were examined by cell counting kit -8 and flow cytometry assay. The targeting relationship was verified by the dual-luciferase reporter and RNA Immunoprecipitation assay.

RESULTS

Highly expressed MIAT and Dicckopf-1 (DDK1), and lowly expressed miR-204-5p were found in the gingival crevicular fluid of periodontitis patients and Pg-LPS induced hPDLFs. MIAT has a sensitivity of 76.53 % and a specificity of 86.49 % for identifying patients with periodontitis among healthy individuals. MIAT acts as a sponge for miR-204-5p and upregulates DDK1 mRNA expression. Silencing of MIAT diminished the promotion of apoptosis and inflammation in hPDLFs by Pg-LPS and enhanced osteogenic differentiation. However, a miR-204-5p inhibitor significantly reversed the effect of silenced MIAT.

CONCLUSIONS

MIAT may act as a promising biomarker for periodontitis. It modulates apoptosis, inflammation, and osteogenic differentiation of PDLFs by focusing on the miR-204-5p/DKK1 axis, indicating its potential as a new therapeutic target for treating periodontitis.

摘要

目的

本研究探讨了长链非编码 RNA 心肌梗死相关转录物(MIAT)在牙周炎中的临床意义及其对人牙周膜成纤维细胞(hPDLFs)功能调节的影响。

方法

纳入 98 例牙周炎患者和 74 例健康对照者。采用牙龈卟啉单胞菌脂多糖(Pg-LPS)刺激 hPDLFs 建立体外细胞模型。实时定量聚合酶链反应检测 MIAT 和成骨因子的 mRNA 水平。酶联免疫吸附试验检测炎症因子浓度。细胞计数试剂盒-8 和流式细胞术检测细胞活力和凋亡。双荧光素酶报告和 RNA 免疫沉淀试验验证靶向关系。

结果

牙周炎患者龈沟液和 Pg-LPS 诱导的 hPDLFs 中 MIAT 和 Dicckopf-1(DDK1)高表达,miR-204-5p 低表达。MIAT 对鉴别牙周炎患者和健康个体的敏感性为 76.53%,特异性为 86.49%。MIAT 作为 miR-204-5p 的海绵体,上调 DDK1 mRNA 表达。沉默 MIAT 可减轻 Pg-LPS 诱导的 hPDLFs 凋亡和炎症,并增强成骨分化。然而,miR-204-5p 抑制剂显著逆转了沉默 MIAT 的作用。

结论

MIAT 可能是牙周炎有前途的生物标志物。它通过靶向 miR-204-5p/DDK1 轴调节 PDLFs 的凋亡、炎症和成骨分化,表明其作为治疗牙周炎的新治疗靶点的潜力。

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