消退素E1调控牙周膜成纤维细胞改善牙周组织炎症的研究
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摘要
目的:探究局部应用消退素E1(RvE1)在小鼠牙周组织炎症中的作用,及其对牙周膜成纤维细胞的影响。方法:C57BL/6小鼠随机分为正常对照组、实验性牙周炎+PBS组和实验性牙周炎+RvE1组,采用结扎法建立实验性牙周炎模型,每2 d局部注射RvE1 1次,10 d后观察牙周组织内炎症细胞浸润状态;分离培养人牙周膜成纤维细胞(hPDLCs),检测牙周组织及hPDLCs中RvE1受体ChemR23的表达情况;以1μg/mL脂多糖模拟体内炎症环境,选取不同浓度梯度的RvE1干预hPDLCs,6、24、72 h后检测炎症因子IL-1β、IL-6、IL-8的表达水平。结果:RvE1的局部注射降低了小鼠牙周组织内炎症浸润程度(P<0.05);ChemR23在正常及炎症牙周组织中广泛表达,分布于hPDLCs胞核周围;1 nmol/L RvE1可抑制hPDLCs中IL-1β、IL-6、IL-8在mRNA和蛋白水平的表达(P<0.05)。结论:RvE1通过调控PDLCs影响炎症微环境中炎症因子的表达水平,进而促进炎症的消退。
Objective: To investigate the effect of local application of resolvin E1(RvE1) on periodontal inflammation in mice and its regulation of periodontal ligament cells. Methods: C57 BL/6 mice were randomly divided into normal group, control group(periodontitis + PBS) and RvE1 group(periodontitis + RvE1). Silk ligation was used to construct periodontitis models on mice, RvE1 was locally injected every other day for 10 days in all. The inflammation changes in periodontium was compared. And the expression of ChemR23 was checked. Human periodontal ligament cells(hPDLCs) were isolated and cultured in vitro. 1 μg/mL lipopolysaccharide was added to culture medium to induce the inflammation condition. Different concentration of RvE1 was added, and then the expression of inflammatory factors, IL-1β, IL-6 and IL-8 was detected. Results: The inflammation infiltration was depressed with the local injections of RvE1. PDLCs express ChemR23 receptor around the nuclei in normal and inflammatory state. 1 nmol/L RvE1 suppressed the expression of IL-1β, IL-6 and IL-8 in periodontal ligament cells at RNA and protein levels. Conclusions: RvE1 inhibits the expression of inflammatory cytokines in periodontal ligament cells and suppresses the inflammatory status of periodontal tissues.
Objective: To investigate the effect of local application of resolvin E1(RvE1) on periodontal inflammation in mice and its regulation of periodontal ligament cells. Methods: C57 BL/6 mice were randomly divided into normal group, control group(periodontitis + PBS) and RvE1 group(periodontitis + RvE1). Silk ligation was used to construct periodontitis models on mice, RvE1 was locally injected every other day for 10 days in all. The inflammation changes in periodontium was compared. And the expression of ChemR23 was checked. Human periodontal ligament cells(hPDLCs) were isolated and cultured in vitro. 1 μg/mL lipopolysaccharide was added to culture medium to induce the inflammation condition. Different concentration of RvE1 was added, and then the expression of inflammatory factors, IL-1β, IL-6 and IL-8 was detected. Results: The inflammation infiltration was depressed with the local injections of RvE1. PDLCs express ChemR23 receptor around the nuclei in normal and inflammatory state. 1 nmol/L RvE1 suppressed the expression of IL-1β, IL-6 and IL-8 in periodontal ligament cells at RNA and protein levels. Conclusions: RvE1 inhibits the expression of inflammatory cytokines in periodontal ligament cells and suppresses the inflammatory status of periodontal tissues.
引文
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[20] Fan C,Zhang X,Upton Z.Anti-inflammatory effects of shikonin in human periodontal ligament cells[J/OL].Pharm Biol,2018,56(1):415-421[2019-03-24].https://doi.org/10.1080/13880209.2018.1506482.
[2] Nicu EA,Loos BG.Polymorphonuclear neutrophils in periodontitis and their possible modulation as a therapeutic approach[J].Periodontol 2000,2016,71(1):140-163.
[3] Serhan CN,Savill J.Resolution of inflammation:the beginning programs the end[J].Nat Immunol,2005,6(12):1191-1197.
[4] Levy BD.Resolvins and protectins:natural pharmacophores for resolution biology[J].Prostaglandins Leukot Essent Fatty Acids,2010,82(4/6):327-332.
[5] Salvi GE,Lang NP.Host response modulation in the management of periodontal diseases[J].J Clin Periodontol,2005,32(Suppl 6):108-129.
[6] Liu G Liu Q,Shen Y,et al.Early treatment with Resolvin E1 facilitates myocardial recovery from ischaemia in mice[J/OL].Br J Pharmacol,2018,175(8):1205-1216[2019-03-24].https://doi.org/10.1111/bph.14041.
[7] Van Dyke TE.Control of inflammation and periodontitis[J].Periodontol 2000,2007,45:158-166.
[8] Fredman G,Oh SF,Ayilavarapu S,et al.Impaired phagocytosis in localized aggressive periodontitis:rescue by Resolvin E1[J/OL].PLoS One,2011,6(9):e24422[2019-03-24].https://doi.org/10.1371/journal.pone.0024422.
[9] Balta MG,Loos BG,Nicu EA.Emerging Concepts in the Resolution of Periodontal Inflammation:A Role for Resolvin E1[J/OL].Front Immunol,2017,8:1682[2019-03-24].https://doi.org/10.3389/fimmu.2017.01682.
[10] Yang X,Zhang H,Wang J,et al.Puerarin decreases bone loss and collagen destruction in rats with ligature-induced periodontitis[J].J Periodontal Res,2015,50(6):748-757.
[11] Naito Y,Okuda K,Takazoe I.Immunoglobulin G response to subgingival gram-negative bacteria in human subjects[J].Infect Immun,1984,45(1):47-51.
[12] Lu Z,Li Y,Brinson CW,et al.CD36 is upregulated in mice with periodontitis and metabolic syndrome and involved in macrophage gene upregulation by palmitate[J/OL].Oral Dis,2017,23(2):210-218[2019-03-24].https://doi.org/10.1111/odi.12596.
[13] Hasturk H,Kantarci A,Ohira T,et al.RvE1 protects from local inflammation and osteoclast- mediated bone destruction in periodontitis[J].FASEB J,2006,20(2):401-403.
[14] Lee CT,Teles R,Kantarci A,et al.Resolvin E1 Reverses Experimental Periodontitis and Dysbiosis[J].J Immunol,2016,197(7):2796-2806.
[15] Wan M,Godson C,Guiry PJ,et al.Leukotriene B4/antimicrobial peptide LL-37 proinflammatory circuits are mediated by BLT1 and FPR2/ALX and are counterregulated by lipoxin A4 and resolvin E1[J].FASEB J,2011,25(5):1697-1705.
[16] Deyama S,Shimoda K,Suzuki H,et al.Resolvin E1/E2 ameliorate lipopolysaccharide-induced depression-like behaviors via ChemR23[J/OL].Psychopharmacology (Berl),2018,235(1):329-336[2019-03-24].https://doi.org/10.1007/s00213-017-4774-7.
[17] Alessi DR,Caudwell FB,Andjelkovic M,et al.Molecular basis for the substrate specificity of protein kinase B;comparison with MAPKAP kinase-1 and p70 S6 kinase[J].FEBS Lett,1996,399(3):333-338.
[18] El-Awady AR,Messer RL,Gamal AY,et al.Periodontal ligament fibroblasts sustain destructive immune modulators of chronic periodontitis[J].J Periodontol,2010,81(9):1324-1335.
[19] Liu J,Tang X,Li C,et al.Porphyromonas gingivalis promotes the cell cycle and inflammatory cytokine production in periodontal ligament fibroblasts[J].Arch Oral Biol,2015,60(8):1153-1161.
[20] Fan C,Zhang X,Upton Z.Anti-inflammatory effects of shikonin in human periodontal ligament cells[J/OL].Pharm Biol,2018,56(1):415-421[2019-03-24].https://doi.org/10.1080/13880209.2018.1506482.