丝氨酸/苏氨酸蛋白激酶促进慢性根尖周炎模型小鼠的骨破坏  

Serine/threonine protein kinases can promote bone destruction in mouse models of chronic periapical periodontitis

作　　者：杨彩会 刘启成[1] 董明[1] 王丽娜[1] 左美娜 陆颖 牛卫东[1] Yang Caihui;Liu Qicheng;Dong Ming;Wang Lina;Zuo Meina;Lu Ying;Niu Weidong(School of Stomatology,Dalian Medical University,Dalian 116044,Liaoning Province,China;Stomatological Hospital,School of Stomatology,Dalian Medical University,Dalian 116000,Liaoning Province,China)

机构地区：[1]大连医科大学口腔医学院,辽宁省大连市116044 [2]大连医科大学口腔医学院附属口腔医院,辽宁省大连市116000

出　　处：《中国组织工程研究》2021年第23期3654-3659,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81700962),项目负责人:陆颖。

摘　　要：背景:根尖周炎的主要表现为骨破坏和形成炎症性肉芽组织,研究发现丝氨酸/苏氨酸蛋白激酶(AKT)可促进破骨细胞分化。目的:观察AKT在小鼠实验性根尖周炎组织中的表达变化。方法:选用25只C57BL/6J野生型雌性小鼠,通过双侧下颌第一磨牙开髓后将髓腔暴露于口腔中的方法,建立小鼠根尖周炎动物模型。将5只未开髓的小鼠作为健康对照组,其余20只作为实验组,开髓后1,2,3,4周每周随机处死5只小鼠,分离下颌骨,制作冰冻切片。采用苏木精-伊红染色,观察小鼠根尖组织炎症情况;采用免疫组织化学染色,观察AKT的表达与分布情况;采用酶组织化学染色,观察根尖组织破骨细胞的表达,并分析AKT与破骨细胞表达的相关性。结果与结论:①苏木精-伊红染色结果显示:健康对照组小鼠根尖周围组织仅见少量炎性细胞,牙周组织完整;开髓术后1周,根尖周组织牙周膜有少量增宽,有小范围的中性粒细胞浸润;开髓术后2周,牙周膜宽度明显增大,根尖周围组织可见大量的中性粒细胞、巨噬细胞、淋巴细胞浸润,出现了明显牙槽骨吸收;开髓术后三四周,炎症范围继续扩大,主要为淋巴细胞浸润,牙槽骨的吸收范围更加明显,牙周膜宽度继续增宽;说明小鼠慢性根尖周炎动物模型建立成功;②AKT免疫组织化学染色结果显示:健康对照组仅有少量阳性细胞表达;实验组AKT的表达量在开髓后1周时开始增多;开髓后2周时达到高峰;三四周时数目下降;各实验组AKT阳性细胞数均高于健康对照组,1周与4周之间相比差异无显著性意义(P>0.05),其余各组间差异均有显著性意义(P<0.05);③酶组织化学染色结果显示:破骨细胞数目随着时间延长而呈现一定的趋势,在开髓后二三周时达到高峰;4周时下降;AKT吸光度值与破骨细胞计数值之间存在中度相关性(r=0.634,P<0.001);④提示AKT在小鼠慢性根尖周炎中有表达,且表达增高;AKBACKGROUND:The main manifestations of periapical periodontitis are bone destruction and formation of inflammatory granulation tissue.Studies have found that serine/threonine protein kinases(AKT)can promote osteoclast differentiation.OBJECTIVE:To observe the expression of AKT in mice with experimental periapical periodontitis.METHODS:Twenty-five C57BL/6J wild-type female mice were selected,and the animal model of periapical periodontitis was established in 20 mice by pulpectomy to expose the pulp cavity of bilateral mandibular first molars.The remaining five mice without pulpectomy were used as the healthy control group.Five mice from the experimental group were randomly killed to separate the mandible and prepare frozen sections at each observational time,including 1,2,3,and 4 weeks after pulpectomy.Hematoxylin-eosin staining was used to observe the inflammation of the mouse apical tissue.Immunohistochemical staining was used to observe the expression and distribution of AKT.Enzyme histochemical staining was used to observe the expression of osteoclasts in the mouse apical tissue.Correlation between AKT and osteoclast expression was analyzed.RESULTS AND CONCLUSION:Hematoxylin-eosin staining results showed that in the healthy control group,only a small amount of inflammatory cells were seen in the tissues around the apex,and the periodontal tissue was intact.At 1 week after the pulpectomy,the periodontal ligament in the periapical tissue was slightly widened,and there were a small amount of infiltrated neutrophils.At 2 weeks after pulpectomy,the width of the periodontal ligament increased significantly,and a large number of neutrophils,macrophages,and lymphocytes infiltrated in the tissues around the apex,and obvious alveolar bone resorption appeared.At 3 to 4 weeks after the pulpectomy,mainly due to lymphocyte infiltration,the range of inflammation continued to expand,the range of alveolar bone resorption was more obvious,and the periodontal ligament width continued to widen.These indicated the successful establi

关 键 词：根尖周炎 AKT 蛋白激酶B 破骨细胞 牙周膜 免疫组织化学 染色 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]