尼古丁对人牙周膜细胞长链非编码RNA NEAT1表达的影响

发布时间：2018-11-10 10:30

【摘要】：目的多种长链非编码RNA可影响人牙周膜细胞的生理反应,而尼古丁作为吸烟相关性牙周炎的主要危险因素,可引起人牙周膜细胞内多种细胞因子的变化。文中旨在探究尼古丁对人牙周膜细胞(h PDLCs)核内长链非编码RNA NEAT1表达水平和白细胞介素(IL)-8表达水平的影响。方法体外培养h PDLCs,第4代细胞用于后续实验。将细胞随机分为对照组、α-BTX组和尼古丁+α-BTX组。对照组:不加任何刺激的人牙周膜细胞;尼古丁组:10-5mol/L尼古丁刺激细胞4 h;α-BTX组:10-8mol/Lα-BTX刺激细胞;尼古丁+α-BTX组:10-5mol/L尼古丁+10-8mol/Lα-BTX共同刺激。qRT-PCR检测细胞NEAT1及下游IL-8 mRNA表达水平;ELISA检测培养基中IL-8蛋白表达情况。结果牙周膜组织块原代培养3周后可见组织块周围细胞成放射状排列,稳定传代后镜下观察可见h PDLCs呈长梭形或纺锤形,轮廓清晰,贴壁生长。与尼古丁组比较,其余3组h PDLCs NEAT1-1、NEAT1-2和IL-8 mRNA表达均降低(P0.05)。尼古丁组培养上清中IL-8蛋白表达水平较对照组显著升高,约为对照组的4.8倍[(1226.0±163.0)pg/m L vs(256.4±49.3)pg/m L,P0.05],而α-BTX组和尼古丁+α-BTX组的IL-8蛋白表达水平较对照组差异无统计学意义(P0.05)。结论尼古丁可促进h PDLCs NEAT1和IL-8的表达,提示NEAT1可能通过促进早期炎性反应参与吸烟相关性牙周炎的发生发展。
[Abstract]:Objective multiple long chain noncoding RNA can affect the physiological response of human periodontal ligament cells. Nicotine, as the main risk factor of smoking associated periodontitis, can cause the change of many cytokines in human periodontal ligament cells. The aim of this study was to investigate the effects of nicotine on the expression of long chain non-coding RNA NEAT1 and interleukin (IL) 8 in (h PDLCs) nucleus of human periodontal ligament cells. Methods the 4 th passage of h PDLCs, cells were cultured in vitro for further experiment. The cells were randomly divided into control group, 伪-BTX group and nicotine-伪-BTX group. Control group: human periodontal ligament cells without any stimulation; nicotine group: 10-5mol/L nicotine stimulated cells for 4 h; 伪-BTX group: 10-8mol/L 伪-BTX stimulated cells; Nicotine 伪-BTX group: 10-5mol/L nicotine 10-8mol/L 伪-BTX was co-stimulated. QRT-PCR was used to detect the expression of NEAT1 and IL-8 mRNA downstream, and ELISA was used to detect the expression of IL-8 protein in culture medium. Results after 3 weeks of primary culture of periodontal ligament tissue mass, the cells around the tissue mass could be seen to be radially arranged. After stable passage, it was observed that h PDLCs was fusiform or spindle-shaped, with clear outline and adherent growth. Compared with nicotine group, the expression of h PDLCs NEAT1-1,NEAT1-2 and IL-8 mRNA in the other three groups were decreased (P0.05). The expression of IL-8 protein in the culture supernatant of nicotine group was significantly higher than that in the control group, which was 4.8-fold of that in the control group [(1226.0 卤163.0) pg/m L vs (256.4 卤49.3 pg/m L (P0.05)]. The expression of IL-8 protein in 伪-BTX group and nicotine 伪-BTX group was not significantly higher than that in control group (P0.05). Conclusion nicotine can promote the expression of h PDLCs NEAT1 and IL-8, suggesting that NEAT1 may be involved in the occurrence and development of smoking-related periodontitis by promoting early inflammatory response.
【作者单位】：军事口腔医学国家重点实验室国家口腔疾病临床医学研究中心陕西省口腔疾病临床医学研究中心第四军医大学口腔医院儿童口腔科;第四军医大学微生物学教研室;
【基金】：国家自然科学基金(81170964,81470743,81670988)
【分类号】：R781.4

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