基于转录组测序探讨柚皮素对脂多糖作用下人牙周膜干细胞的抗炎作用及相关机制
Anti-inflammatory effects and related mechanisms of naringenin in human periodontal ligament stem cells under lipopolysaccharide stimulation based on RNA sequencing
作者：李俊玉, 徐晓梅, 刘兴玉, 曾婷, 张丽, 郑茜
Author：Li Junyu, Xu Xiaomei, Liu Xingyu, Zeng Ting, Zhang Li, Zheng Qian
收稿日期：2023-12-26 年卷（期）页码：2024,42(4):512-512-520
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：柚皮素,人牙周膜干细胞,抗炎作用,核因子κB,转录组测序,
Key words：naringenin,human periodontal ligament stem cells,anti-inflammatory effect,nuclear factor κB,RNA sequencing,
基金项目：四川省科技厅创新专项项目(2022YFS0634);四川省中医药管理局面上项目(2023MS080);泸州市科技计划项目(2022-JYJ-116)
中文摘要
目的 联合使用转录组测序（RNA-seq）和生信分析，探究柚皮素（Nar）对脂多糖（LPS）刺激下的人牙周膜干细胞（hPDLSCs）的抗炎作用及机制。 方法 采用细胞计数试剂盒（CCK-8）、实时荧光定量逆转录聚合酶链反应（qRT-PCR）、酶联免疫吸附试验（ELISA）检测Nar对LPS刺激下hPDLSCs增殖及炎性因子表达情况，筛选出Nar的最佳抗炎浓度。采用RNA-seq，以|log2FC|≥1且P≤0.05为标准筛选出显著差异基因（DEGs）。采用火山图分析、京都基因与基因组百科全书（KEGG）通路富集分析、String数据库及Cytoscape的MCODE模块筛选核心基因。ELISA、qRT-PCR和蛋白印迹实验（Western blot）检测对核因子κB（NF-κB）信号通路的影响。 结果 适宜浓度的Nar可减轻LPS刺激下hPDLSCs的炎症因子表达，促进其增殖，20 μmol/L Nar抗炎效果最佳。RNA-seq提示炎症相关信号通路显著富集。Nar通过抑制NF-κB信号通路产生抗炎作用，Nar与NF-κB抑制剂BAY 11-7802效果相似。 结论 Nar可以通过抑制NF-κB信号通路发挥抗炎作用。Nar可能是辅助治疗牙周炎的一种潜在的药效成分。
英文摘要
ObjectiveRNA sequencing (RNA-seq) and bioinformatic analysis were combined and used to explore the anti-inflammatory effects and mechanisms of naringenin (Nar) in lipopolysaccharide (LPS)-stimulated human periodontal ligament stem cells (hPDLSCs).MethodsCell counting kit-8, quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), and enzyme-linked immunosorbent assay (ELISA) were adopted to detect the effects of Nar on the proliferation and expression of inflammatory factors in LPS-stimulated hPDLSCs, screening for the optimal anti-inflammatory concentration of Nar. Differentially expressed genes (DEGs) were screened using |log2FC|≥1 andP≤0.05 as criteria. Volcano plot analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, the String database, and the MCODE module of Cytoscape were utilized to select core genes and enriched pathways. The effects on the nuclear factor κB (NF-κB) signaling pathway were verified using ELISA, qRT-PCR, and Western blot.ResultsAppropriate concentrations of Nar could alleviate the expression of inflammatory factors and promote the proliferation of hPDLSCs stimulated by LPS. The best anti-inflammatory effect was achieved with 20 μmol/L Nar. RNA-seq showed significant enrichment of inflammation-related signaling pathways. The anti-inflammatory effect of Nar was mediated by inhibiting the NF-κB signaling pathway, similar to the effect of the NF-κB inhibitor BAY 11-7802.ConclusionNar could exert its anti-inflammatory effects by inhibiting the NF-κB signaling pathway, making it a potential therapeutic option for the adjuvant treatment of periodontitis.
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