摘要
目的:探讨炎性因子刺激下免疫调控因子程序性死亡因子配体1(PD-L1)在人牙周膜成纤维细胞表面的表达情况。方法:体外培养人牙周膜成纤维细胞和牙周致病菌。分别用0,5,10,20 ng/mL的浓度梯度的TNF-α和IFN-γ诱导牙周膜成纤维细胞,在0,24,48,72,96 h5个时间点,收集牙周膜成纤维细胞,分析PD-L1在牙周膜成纤维细胞表面表达的时间和剂量依赖性。牙龈卟啉单胞菌的培养上清和活化后的单核细胞培养上清诱导牙周膜成纤维细胞,利用流式细胞仪分析其表面PD-L1的表达。炎性因子IL-1、IL-6、TNF-α、IFN-γ、LPS及上述炎性因子的混合物(combination)和牙周致病菌卟啉单胞菌、中间普雷沃菌和具核梭杆菌来诱导牙周膜成纤维细胞表面,利用Western Blot技术和qPCR检测PD-L1的表达情况。结果:TNF-α和IFN-γ诱导牙周膜成纤维细胞,PD-L1在牙周膜成纤维细胞表面的表达具有时间和剂量依赖性。Western Blot技术和qPCR也验证了炎性因素刺激可以上调牙周膜成纤维细胞表面PD-L1的表达。结论:炎性刺激下牙周膜成纤维细胞PD-L1表达水平显著上调,且TNF-α和IFN-γ刺激下PD-L1的表达具有时间和剂量的依赖性。PD-L1的表达可能与牙周炎有相关关系。
Objective:To explore the programmed death 1 ligand 1(PD-L1)expression induced by proinflammatory factors on the surface of periodontal ligament cells in vitro.Methods:Periodontal ligament cells were isolated from periodontal ligament tissues of premolar teeth of three donors and cultured in vitro.Periodontal pathogens,Porphyromonas gingivalis,Prevotella intermedia,and Fusobacterium nucleatum were obtained from State Key Laboratory of Oral Diseases(Chengdu,China)and cultured in brain heart infusion(BHIB)broth under anaerobic conditions at 37℃for 48 h.PD-L1 expression on the surface of PDLCs was induced respectively by TNF-α,IFN-γ,LPS,and periodontal pathogens,and was analyzed by flow cytometry,Western Blot,and immunohistochemistry,respectively.Results:The expression of PD-L1 on the surface of periodontal ligament cells induced by TNF-αand IFN-γwas in a time-and dose-dependent manner,respectively.PD-L1 was inducibly expressed on PDLCs stimulated by inflammatory cytokines and periodontal pathogens through confirmed detection by Western Blot and qPCR.Conclusion:PD-L1 is inducibly expressed on PDLCs stimulated by inflammatory cytokines and periodontal pathogens.
作者
张杰华
张平
李伟
李俊
彭友俭
ZHANG Jiehua;ZHANG Ping;LI Wei;LI Jun;PENG Youjian(Dept.of Stomatology,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei,China;State Key Laboratory of Oral Diseases,West China Hospital of Stomatology of Sichuan University,Chengdu 610000,Sichuan,China)
出处
《武汉大学学报（医学版）》
CAS
2019年第6期871-876,共6页
Medical Journal of Wuhan University
基金
武汉大学自主科研项目(编号:2042017kf0166).
关键词
牙周膜成纤维细胞
程序性死亡因子配体1
炎性因子
Periodontal Ligament Cells
Programmed Cell Death 1 Ligand 1
Inflammatory Cytokine
