敲除巨噬细胞Zfp260通过抑制M1型极化减轻小鼠牙周炎症  

Knockout of macrophage Zfp260 attenuates periodontitis in mice via inhibiting M1-type polarization

作　　者：熊瑾 王佐林[1] XIONG Jin;WANG Zuolin(Shanghai Engineering Research Center of Tooth Restoration and Regeneration&Tongji Research Institute of Stomatology&Department of Oral Implantology,Stomatological Hospital and Dental School,Tongji University,Shanghai 200072,China)

机构地区：[1]同济大学附属口腔医院口腔种植科,同济大学口腔医学院,上海牙组织修复与再生工程技术研究中心,同济大学口腔医学研究所,上海200072

出　　处：《口腔颌面外科杂志》2024年第4期259-267,共9页Journal of Oral and Maxillofacial Surgery

基　　金：科技部重点研发专项(2018YFE0202200);国家自然科学基金(81670962)。

摘　　要：目的:研究锌指蛋白260(zinc finger protein 260,Zfp260/ZNF260)对巨噬细胞极化和牙周炎牙槽骨吸收的影响。方法:实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测人/野生型小鼠炎症牙周组织中ZNF260/Zfp260的表达。RT-qPCR检测RAW264.7细胞在炎症和非炎症环境中Zfp260的表达。繁育巨噬细胞内特异性敲除Zfp260小鼠,分别构建flox/flox对照鼠(Zfp260^(f/f);Lyz2-cre^(-))和条件性敲除(conditional knockout,cKO)鼠(Zfp260^(f/f);Lyz2-cre^(+))牙周炎模型,micro-CT、苏木精-伊红(hematoxylin and eosin,HE)染色观察牙槽骨形态变化。RT-qPCR、蛋白质印迹法观察应用特异性小干扰RNA(small interfering RNA,siRNA)沉默Zfp260表达对RAW264.7极化的影响。RT-qPCR、免疫荧光染色观察牙周组织中M1、M2型极化标志分子的表达。结果:人/野生型小鼠炎症牙周组织中ZNF260/Zfp260的表达明显高于健康组。炎症环境下RAW264.7细胞中Zfp260的表达明显上调。cKO小鼠的牙周炎侧骨吸收小于flox/flox小鼠。沉默Zfp260表达可抑制RAW264.7在炎症环境下的M1型极化,且cKO小鼠的炎症牙周组织中M1型极化标志分子的表达明显少于flox/flox小鼠。结论:干扰巨噬细胞Zfp260的表达可抑制M1型极化并减轻牙周炎导致的牙槽骨吸收。Objective:To explore the effect of zinc finger protein 260(Zfp260/ZNF260)on macrophage polarization and alveolar bone resorption due to periodontitis.Methods:Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of ZNF260/Zfp260 in inflammatory periodontal tissues of human/wild-type mice.The expression of Zfp260 in RAW264.7 cells in inflammatory or non-inflammatory environments was detected by RT-qPCR.Mice with specific knockout of Zfp260 were bred,and control mice(Zfp260^(f/f);Lyz2-cre^(-))and conditional knockout(cKO)mice(Zfp260^(f/f);Lyz2-cre^(+))periodontitis models were established respectively.Morphological changes of alveolar bone were detected by micro-CT and hematoxylin-eosin(HE)staining.The polarization of RAW264.7 cells when Zfp260 was knocked down via specific small interfering RNA(siRNA)in an inflammatory environment was detected by RT-qPCR and western blotting.RT-qPCR and immunofluorescence staining were used to observe the expression of M1 and M2 macrophage-associated markers in periodontal tissues.Results:The expression of ZNF260/Zfp260 in inflammatory periodontal tissues of human/wild-type mice were significantly higher than that of healthy periodontal tissues.The expression of Zfp260 in RAW264.7 cells was significantly increased in an inflammatory environment.Alveolar bone resorption in the ligatured side of cKO mice was significantly less than that of flox/flox mice.The knockdown of Zfp260 in RAW264.7 cells could inhibit its M1 polarization in an inflammatory environment.The expression of M1-related markers in inflammatory periodontal tissues was significantly lower than that of flox/flox mice.Conclusion:Inhibition of Zfp260 in macrophage decreased M1 polarization and rescued the bone loss due to periodontitis.

关 键 词：锌指蛋白 牙周炎 巨噬细胞 极化 

分 类 号：R782[医药卫生—口腔医学]