静压力下牙周膜成纤维细胞分泌炎性因子与p38信号通路的关系  

Relationship between p38 signaling pathway and periodontal ligament fibroblasts secreting inflammatory factors under static pressure

作　　者：唐海芳 彭娟敏 康娜[1,2,3,4] Tang Haifang;Peng Juanmin;Kang Na(Hospital of Stomatology,Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China;Key Laboratory of Oral and Maxillofacial Restoration and Reconstruction of Guangxi Zhuang Autonomous Region,Nanning 530021,Guangxi Zhuang Autonomous Region,China;Craniomaxillofacial Clinical Research Center of Guangxi Zhuang Autonomous Region,Nanning 530021,Guangxi Zhuang Autonomous Region,China;Key Laboratory of Diagnosis and Treatment of Maxillofacial Surgery (Key Laboratory of Guangxi University),Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学附属口腔医院,广西壮族自治区南宁市530021 [2]广西口腔颌面修复与重建研究自治区级重点实验室,广西壮族自治区南宁市530021 [3]广西颅颌面畸形临床医学研究中心,广西壮族自治区南宁市530021 [4]颌面外科疾病诊治研究重点实验室(广西高校重点实验室),广西壮族自治区南宁市530021

出　　处：《中国组织工程研究》2019年第15期2308-2313,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81360170);项目负责人:康娜~~

摘　　要：背景:研究表明p38 MAPK通路在炎症因子的合成中起到重要作用,但对正畸牙移动过程牙周膜成纤维细胞中p38 MAPK信号通路与白细胞介素17、白细胞介素6等炎性因子关系的相关报道较少。目的:分析人牙周膜成纤维细胞加载持续性静压力后在合成炎症因子白细胞介素6、白细胞介素17过程与p38MAPK的关系,探讨p38 MAPK信号通路对白细胞介素17和白细胞介素6表达的影响。方法:①取年龄在11-15岁正畸者需要拔除的新鲜第一、二前磨牙的牙周组织(正畸者及其监护人均知情同意),体外培养牙周膜成纤维细胞,建立细胞重力加载模型;②细胞种板,分别加力0,1,2,4 g/cm^2,每加力组分别加力5,15,30和60min,检测p38MAPK蛋白表达;③取第4代细胞接种后,随机分为6组:空白组、加二甲基亚砜组、加抑制剂组(p38 MAPK阻断剂SB203580)、加力组、加力+二甲基亚砜组、加力+抑制剂组,对细胞预处理60 min,通过定量RT-qPCR和酶联免疫吸附法(ELISA)检测牙周膜成纤维细胞受4 g/cm^2压力刺激后白细胞介素17和白细胞介素6的mR NA和蛋白水平。结果与结论:①2 g/cm^2组、4 g/cm^2组p38 MAPK磷酸化蛋白表达变化随时间的延长而增加:30 min时表达量最高,60min时开始减少;②与未加力的3组相比,加力组的白细胞介素17和白细胞介素6表达量上调,加抑制剂组的白细胞介素6表达量下降,白细胞介素17表达量无明显差异;③结果提示,静压力可促使牙周膜成纤维细胞分泌白细胞介素17和白细胞介素6,且白细胞介素6的分泌可能与p38MAPK信号通路有关,但不足以证明该通路参与静压力对牙周膜成纤维细胞诱导白细胞介素17的表达。BACKGROUND:p38 MAPK pathway has been shown to play an important role in the synthesis of inflammatory factors,but the relationship between p38 MAPK signaling pathway and interleukin 17,interleukin 6 and other inflammatory factors in periodontal ligament fibroblasts during orthodontic tooth movement is little reported.OBJECTIVE:To investigate the relationship of the expression of inflammatory cytokines interleukin 17 and interleukin 6 with p38 MAPK in human periodontal ligament fibroblasts under continuous static pressure,and to explore the effect of p38 MAPK signaling pathway on the expression of interleukin 17 and interleukin 6.METHODS:The periodontal tissues of fresh first and second premolars were obtained from the removed teeth of the orthodontic patients aged 11-15 years(informed consent had been obtained).Periodontal ligament fibroblasts were cultured in vitro,and the cell model loaded with gravity was established.Cell culture plates were loaded with 0,1,2 and 4 g/cm^2 pressure for 5,15,30 and 60 minutes,respectively.The expression of p38 MAPK protein was detected.The passage 4 cells were seeded,and then randomized into six groups:blank,dimethyl sulfoxide,inhibitor(p38 MAPK blocker,SB203580),loading,loading plus dimethyl sulfoxide,and loading plus inhibitor groups.The intervention lasted for 60 minutes.The interleukin 17 and interleukin 6 protein and mRNA levels in periodontal ligament fibroblasts after loaded with 4 g/cm^2 force were detected by RT-PCR and ELISA.RESULTS and CONCLUSION:(1)The expression level of p38 MAPK protein in the 2 and 4 g/cm^2 groups was increased with time prolonged,peaked at 30 minutes,and began to decrease at 60 minutes.(2)Compared with the groups without loading,the loading groups showed an increase in the expression levels of interleukin 17 and interleukin 6.The expression level of interleukin 6 in the inhibitor group was down-regulated,and the expression level of interleukin 17 showed no significant difference.(3)To conclude,static pressure can promote human periodontal ligamen

关 键 词：静压力 牙周膜成纤维细胞 p38 丝裂原激活蛋白 白细胞介素17 白细胞介素6 牙周膜 正畸学 牙移动 应力 物理 组织工程 

分 类 号：R496[医药卫生—康复医学]