微小RNA-218-5p靶向TLR2基因对LPS所致人牙周膜干细胞炎症反应的影响

doi:10.13701/j.cnki.kqyxyj.2020.12.007

摘要/Abstract

摘要： 目的:研究微小RNA(microRNA,miR)-218-5p对脂多糖(LPS)诱导的牙周膜干细胞(PDLSCs)增殖、凋亡和炎症的影响及作用机制。方法:分离培养PDLSCs,用LPS处理PDLSCs细胞建立炎症模型,MTT法和流式细胞术分别检测细胞存活率和凋亡率,qRT-PCR和Western blot分别检测miR-218-5p和Toll样受体2(TLR2)蛋白的表达,检测细胞中白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-ɑ(TNF-ɑ)的含量,双荧光素酶报告系统检测miR-218-5p和TLR2的靶向关系。结果:LPS处理人PDLSCs 0、12、24和48 h,PDLSCs细胞中miR-218-5p表达下调,TLR2表达上调,细胞存活率下降,凋亡率升高,炎症因子IL-6、IL-1β和TNF-α含量升高,呈时间依赖趋势,差异均有统计学意义(P<0.05);miR-218-5p靶向调控TLR2表达;过表达miR-218-5p或沉默TLR2均可提高细胞存活率,降低细胞凋亡率和炎症因子IL-6、IL-1β、TNF-α的含量;过表达TLR2可逆转miR-218-5p过表达对LPS诱导的PDLSCs细胞炎症和细胞增殖的作用。结论:miR-218-5p通过靶向TLR2调控LPS诱导的PDLSCs细胞增殖、凋亡和炎症反应。

关键词: 微小RNA-218-5p, 脂多糖, Toll样受体2, 牙周膜干细胞, 炎症, 存活率, 凋亡

Abstract: Objective: To investigate the effects and mechanism of miR-218-5p on proliferation, apoptosis, and inflammation of periodontal ligament stem cells (PDLSCs) induced by lipopolysaccharide (LPS). Methods: PDLSCs were isolated and cultured. PDLSCs cells were treated with LPS to establish an inflammatory model. MTT assay and flow cytometry were applied to detect the cell survival rate and apoptosis rate, respectively. qRT-PCR and Western blot were performed to detect the expression levels of miR-218-5p and TLR2 protein, respectively. The kits was used to detect the contents of interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in PDLSCs. The dual luciferase reporting system was used to detect the targeting relationship between miR-218-5p and TLR2. Results: Human PDLSs were treated by LPS for 0h, 12h, 24h, and 48h. miR-218-5p in PDLSs cells was down-regulated, TLR2 was up-regulated, cell survival rate was down-regulated, apoptosis rate was up-regulated, and the contents of inflammatory factors IL-6, IL-1β, and TNF-α were up-regulated, showing a time-dependent trend with statistical significance (P<0.05). MiR-218-5p targeted and regulated the expression of TLR2. Over-expression of miR-218-5p or silencing of TLR2 both improved cell survival rate and reduced cell apoptosis rate and levels of inflammatory factors IL-6, IL-1β, and TNF-α. Over-expression of TLR2 reversed the effect of miR-218-5p over-expression on inflammation and proliferation of PDLSCs induced by LPS. Conclusion: MiR-218-5p regulates the proliferation, apoptosis, and inflammatory response of PDLSCs cells induced by LPS through targeting TLR2.

Key words: microRNA-218-5p, lipopolysaccharide, Toll-like receptor 2, periodontal membrane stem cells, inflammation, survival rate, apoptosis

周洋. 微小RNA-218-5p靶向TLR2基因对LPS所致人牙周膜干细胞炎症反应的影响[J]. 口腔医学研究, 2020, 36(12): 1108-1112.

ZHOU Yang. Effect of MicroRNA-218-5p on Inflammation of Human Periodontal Ligament Stem Cells Induced by Lipopolysaccharide through Targeting TLR2 Gene[J]. Journal of Oral Science Research, 2020, 36(12): 1108-1112.

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