Effects of Foxp3 gene silencing on the expression of inflammatory cytokines and the proliferation and migration of human periodontal ligament fibroblasts in an inflammatory environment.

Q3 Medicine
Ting Lu, Jiahao Zhu, Shihe Yang, Zhe Shen, Liangjun Zhong
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Abstract

Objectives: This study aimed to clarify the effects of Foxp3 silencing on the expression of inflammatory cytokines in human periodontal ligament cells (hPDLFs) in an inflammatory environment and on cell proliferation and invasiveness, as well as to explore the role of Foxp3 gene in the development of periodontitis.

Methods: An small interfering RNA (siRNA) construct specific for Foxp3 was transfected into hPDLFs. Foxp3 silencing efficiency was verified by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, and the siRNA with the optimum silencing effect of Foxp3 gene was screened. Using lipopolysaccharide to simulate an inflammatory environment in vitro, CCK-8 detected the effect of silencing Foxp3 on hPDLFs proliferation under inflammatory conditions. Wound-healing experiments and transwell assays were conducted to detect the effect of silencing Foxp3 on hPDLF migration under inflammatory conditions. The expression of the inflammatory cytokines interleukin (IL)-6 and IL-8 was detected by RT-PCR and Western blotting under inflammatory conditions.

Results: After siRNA transfection, RT-PCR and Western blotting analyses showed that the expression of Foxp3 mRNA in the Foxp3-si3 group decreased significantly (t=21.03, P<0.000 1), and the protein expression of Foxp3 also decreased significantly (t=12.8, P<0.001). In the inflammatory environment, Foxp3 gene silencing had no significant effect on hPDLFs proliferation (P>0.05), and Foxp3 gene silencing promoted hPDLFs migration (P<0.05). Moreover, the expression of IL-6 and IL-8 increased (P<0.05).

Conclusions: In an inflammatory environment, Foxp3 gene silencing promoted hPDLFs migration but had no significant effect on hPDLFs proliferation. The expression of inflammatory factors expressed in hPDLFs increased after Foxp3 gene silencing, indicating that Foxp3 gene inhibited inflammation in periodontitis.

Foxp3基因沉默对炎症细胞因子表达及炎症环境下牙周韧带成纤维细胞增殖和迁移的影响
目的:本研究旨在阐明炎症环境下Foxp3沉默对人牙周韧带细胞(hPDLFs)炎症因子表达的影响,以及Foxp3基因对细胞增殖和侵袭性的影响,探讨Foxp3基因在牙周炎发生中的作用。方法:将Foxp3特异性小干扰RNA (siRNA)转染到hPDLFs中。通过逆转录聚合酶链反应(RT-PCR)和Western blotting验证Foxp3的沉默效率,筛选出Foxp3基因沉默效果最佳的siRNA。CCK-8利用脂多糖模拟体外炎症环境,检测炎症条件下沉默Foxp3对hPDLFs增殖的影响。通过创面愈合实验和transwell实验检测炎症条件下沉默Foxp3对hPDLF迁移的影响。采用RT-PCR和Western blotting检测炎症条件下炎性细胞因子IL -6、IL-8的表达。结果:siRNA转染后,RT-PCR和Western blotting分析显示,Foxp3-si3组Foxp3 mRNA表达显著降低(t=21.03, Pt=12.8, PP>0.05), Foxp3基因沉默促进hPDLFs迁移(pp结论:在炎症环境下,Foxp3基因沉默促进hPDLFs迁移,但对hPDLFs增殖无显著影响。Foxp3基因沉默后,hPDLFs中表达的炎症因子表达增加,表明Foxp3基因抑制了牙周炎的炎症。
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来源期刊
华西口腔医学杂志
华西口腔医学杂志 Medicine-Medicine (all)
CiteScore
0.80
自引率
0.00%
发文量
6397
期刊介绍: West China Journal of Stomatology (WCJS, pISSN 1000-1182, eISSN 2618-0456, CN 51-1169/R), published bimonthly, is a peer-reviewed Open Access journal, hosted by Sichuan university and Ministry of Education of the People's Republic of China. WCJS was established in 1983 and indexed in Medline/Pubmed, SCOPUS, EBSCO, Chemical Abstract(CA), CNKI, WANFANG Data, etc.
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