Huang Pengcheng, Jia Linghui
Fujian Key Laboratory of Oral Diseases & Fujian Provincial Engineering Research Center of Oral Biomaterial & Stomatological Key Laboratory of Fujian College and University, School and Hospital of Stomatology, Fujian Medical University, Fuzhou, China.
J Dent Sci. 2022 Oct;17(4):1501-1509. doi: 10.1016/j.jds.2022.04.031. Epub 2022 May 14.
BACKGROUND/PURPOSE: Recent studies have pointed to the crucial role of microRNAs (miRNAs) in chronic periodontitis (CP). This study investigated the regulation and potential mechanisms of miR-28-5p in CP patients and lipopolysaccharide (LPS)-induced periodontal ligament cells (PDLCs).
76 CP patients and 71 periodontally healthy subjects were included. RT-qPCR was employed to examine miR-28-5p and sphingosine kinase -1 (SPHK1) in subjects' gingival sulcus fluid and PDLCs. The diagnostic performance was evaluated by measuring the area under the curve (AUC) of the receiver operating characteristic (ROC) analysis. Pearson correlation coefficient (r) was adopted to explore the statistical relation between indicators. PDLCs proliferation and inflammation factors were determined by CCK-8 and ELISA assay. The direct target gene was validated by a dual-luciferase reporter assay.
miR-28-5p was lowly expressed in CP patients and LPS-induced PDLCs ( < 0.05). AUC for miR-28-5p was 0.937, which had certain diagnostic value. Additionally, miR-28-5p was negatively correlated with periodontal clinical indicators and inflammatory factors. Cell proliferation of PDLCs was inhibited and inflammation was promoted under LPS induction, however, elevated miR-28-5p diminished the effect of LPS ( < 0.05). SPHK1 acts as a miR-28-5p target and the elevation of SPHK1 caused by LPS treatment was inhibited by the increased miR-28-5p.
Present study revealed that miR-28-5p could be served as a potential diagnostic biomarker for CP. And miR-28-5p may participate in CP progression by targeting SPHK1 to regulate the proliferation and inflammation of PDLCs. This study may offer insights into CP treatment and diagnosis.
背景/目的：最近的研究指出微小RNA（miRNA）在慢性牙周炎（CP）中起关键作用。本研究调查了miR-28-5p在CP患者和脂多糖（LPS）诱导的牙周膜细胞（PDLCs）中的调控作用及潜在机制。
纳入76例CP患者和71例牙周健康受试者。采用逆转录定量聚合酶链反应（RT-qPCR）检测受试者龈沟液和PDLCs中miR-28-5p和鞘氨醇激酶-1（SPHK1）的表达。通过测量受试者工作特征（ROC）分析的曲线下面积（AUC）评估诊断性能。采用Pearson相关系数（r）探讨指标间的统计关系。通过细胞计数试剂盒-8（CCK-8）和酶联免疫吸附测定（ELISA）法检测PDLCs增殖和炎症因子。通过双荧光素酶报告基因检测验证直接靶基因。
miR-28-5p在CP患者和LPS诱导的PDLCs中低表达（P<0.05）。miR-28-5p的AUC为0.937，具有一定诊断价值。此外，miR-28-5p与牙周临床指标和炎症因子呈负相关。LPS诱导下PDLCs的细胞增殖受到抑制，炎症反应增强，然而，miR-28-5p表达升高可减弱LPS的作用（P<0.05）。SPHK1是miR-28-5p的靶基因，miR-28-5p表达增加可抑制LPS处理引起的SPHK1表达升高。
本研究表明miR-28-5p可作为CP的潜在诊断生物标志物。并且miR-28-5p可能通过靶向SPHK1调节PDLCs的增殖和炎症反应参与CP的进展。本研究可能为CP的治疗和诊断提供思路。
