辅助性T细胞1和17细胞特征性分泌因子在大鼠实验性牙周炎模型中的表达及意义  

Effects of T helper 1 cells and T helper 17 cells secreting cytokines on rat models of experimental periodontitis

作　　者：王中秀 杨岚[1] 谭静怡[1] 陈莉丽[1] Wang Zhongxiu;Yang Lan;Tan Jingyi;Chen Lili(Department of Periodontology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310009, China)

机构地区：[1]浙江大学医学院附属第二医院牙周病专科,杭州,310009

出　　处：《中华口腔医学杂志》2017年第12期740-746,共7页Chinese Journal of Stomatology

基　　金：国家自然科学基金（81271142、81771072）

摘　　要：目的 探究辅助性T细胞1和17特征性分泌因子干扰素γ和白细胞介素17（interleukin-17,IL-17）在实验性大鼠牙周炎外周血和牙槽骨破坏区域的表达差异,为细胞免疫介导的牙槽骨破坏提供理论依据.方法 选择8周龄健康雄性SD大鼠18只,按随机数字表法随机分为3组：大肠杆菌内毒素脂多糖（lipopolysaccharide,LPS）组、结扎组和正常对照组,每组6只.LPS组大鼠采用上颌第一磨牙颊侧隔天注射LPS的方法建立实验性牙周炎模型,共注射4次;结扎组大鼠采用0.2 mm正畸丝结扎上颌第一磨牙并辅以高糖饮食;正常对照组大鼠不做任何处理,正常喂养.分别于注射后第4周、结扎后第8周处死动物,采用酶联免疫吸附测定法检测大鼠外周血清中干扰素γ和IL-17的含量;HE染色观察各组大鼠牙周组织变化并于光镜下进行破骨细胞计数;抗酒石酸酸性磷酸酶（tartrate-resistant acid phosphatase,TRAP）染色观察牙槽骨组织内破骨细胞情况;免疫组化法检测牙周局部组织中干扰素γ和IL-17的表达.结果 LPS组和结扎组大鼠外周血清干扰素-γ的表达[分别为（185.0±50.7）、（202.9±60.4） ng/L]均显著高于对照组[（106.3±17.2） ng/L] （P＜0.05）;HE染色可见LPS组和结扎组大鼠的牙龈上皮内有不同程度的炎性细胞浸润,牙槽骨高度降低,出现骨吸收陷窝,且LPS组和结扎组破骨细胞计数[分别为（9.05±1.05）、（10.83±1.17）个]均较对照组[（0.33±0.52）个]显著增加（P＜0.05）;TRAP染色显示LPS组和结扎组可见大量体积较大、胞质红染、内有蓝色核仁的多核破骨细胞;此外,LPS组和结扎组大鼠牙槽骨吸收区域IL-17的表达显著高于对照组.结论 采用LPS定点注射法可建立大鼠牙槽骨吸收模型,结扎法可成功建立大鼠实验性牙周炎模型;干扰素γ和IL-17均参与牙周炎发生过程,IL-17可能与牙槽骨吸收破坏有关.Objectvie To investigate the effects of secreting cytokines interferon-gamma （IFN-γ） and interleukin-17 （IL-17） of T helper 1 cells （Th1） and T helper 17 cells （Th17） on the peripheral blood and alveolar bone destruction,so as to provide a new explanation for cellular immunity-mediated alveolar bone destruction.Methods Eighteen eight-week-old male Sprague-Dawley rats were divided,randomly and equally,into 3 groups：lipopolysaccharide （LPS） group,ligation group and normal control group.In the LPS group,Escherichia coli LPS was injected into the alveolar mucosa on the buccalmedian site of the left upper first molar,while the right upper first molar was injected with equal volume of physiological saline as self-controls.The injections were performed every other day for four times totally.In the ligation group,the left upper first molars were ligatured with 0.2 mm orthodontic cords,while the right upper first molars were left untreated as self-controls,and supplemented with high-sugar diet to promote the periodontitis status.The rats in normal control group were fed normally.The concentrations of IFN-γand IL-17 in peripheral blood were measured using enzyme linked immunosorbent assay （ELISA） method at the fourth week after the start of injection and at the eighth week after ligation.The histological of periodontal tissues were observed after hematoxylin-eosin （HE） staining and osteoclast count was performed under light microscope.The histological of osteoclasts were observed after tartrate-resistant acid phosphatase （TRAP） staining.Expression of IFN-γ and IL-17 were detected by immunohistochemical assay.Results The concentrations of IFN-γ in peripheral blood of LPS group [（185.0±50.7） ng/L] and ligation group [（202.9±60.4） ng/L] were significantly higher than that of normal control group [（106.3 ± 17.2） ng/L] （P〈0.05）.Meanwhile,histological examination showed inflammatory cells infiltration in the gingival epithelium,the height reduction of alveolar bone accom

关 键 词：动物 实验 牙周炎 干扰素Γ 白细胞介素17 

分 类 号：R-332[医药卫生] R781.42