小鼠正畸牙移动及牙周组织改建中Sema3A的作用  

The effect of Sema3A on the periodontal tissue remodeling during orthodontic tooth movement of mice

作　　者：陈钰文[1] 白秋野 胡丽华[1] 沈丹阳[1] 阎秀林[1] CHEN Yuwen;BAI Qiuye;HU Lihua;SHEN Danyang;YAN Xiulin(Department of Orthodontics,School of Stomatology,China Medical University,Shenyang 110002,China)

机构地区：[1]中国医科大学口腔医学院正畸一科/辽宁省口腔医学研究所口腔正畸学教研室/辽宁省口腔疾病转化医学研究中心/中国医科大学口腔医学院中心实验室,沈阳110002

出　　处：《疑难病杂志》2018年第4期395-399,404,共6页Chinese Journal of Difficult and Complicated Cases

基　　金：辽宁省教育厅科学技术计划项目(LK201607)

摘　　要：目的探讨在小鼠正畸牙移动以及牙周组织改建中Sema3A的表达情况,明确在此过程中Sema3A表达变化的特定规律性。方法实验于2014年1月在中国医科大学动物实验中心进行,取27只wildtype(C57BL/6)小鼠建立小鼠正畸牙移动模型。上颌左侧第一磨牙与切牙间施加10~15 g力为加力组,上颌右侧为对照组,在第1天、7天、14天分别处死9只wildtype小鼠,3只用于制作石蜡组织切片,应用免疫组织化学染色法和HE染色法观察组织形态变化,3只制作硬组织不脱钙切片,用于钙黄绿素沉积情况并用于Masson染色,3只利用Western-blotting分析Sema3A蛋白表达水平检测。结果所有实验鼠均出现牙齿移动,实验建模成功。(1)对照组牙周组织形态正常,未见骨吸收、骨形成等变化,且成骨细胞和胶原纤维极为少见;加力组在第1天其牙周膜纤维排列和成骨细胞数目未见明显改变,但是胶原纤维数目明显多于对照组;在实验第7天牙周膜纤维排列出现明显紊乱,张力侧可见牙周膜变宽以及大量成骨细胞,并且胶原纤维分布水平显著升高;但是在第14天其组织形态基本恢复正常,成骨细胞和胶原纤维稀少分布。(2)免疫组织化学染色显示,对照组Sema3A始终为阴性表达,加力组第1天Sema3A为弱阳性表达,第7天升至强阳性表达,在第14天恢复至弱阳性表达。(3)Western-blotting检测:2组均出现Sema3A蛋白印迹,定量分析显示对照组Sema3A蛋白未见波动,为低水平表达;加力组第1天Sema3A蛋白表达为低水平;第7天Sema3A蛋白表达量显著升高,其表达水平显著高于对照组(P<0.05);第14天,Sema3A蛋白表达量明显下降,恢复到与对照组水平相近(P>0.05)。结论在小鼠正畸牙移动中,Sema3A有可能参与了相关牙周组织改建,其表达变化具有一定的规律性。在正畸牙移动骨改建过程中,Sema3A有可能促进成骨,对牙槽骨改建可能具有积极的保护作用。Objective To discuss the expression of Sema3A in orthodontic tooth movement and periodontal tissue remodeling in mice,and clarify the specific temporal and spatial regularity of Sema3A expression during the process.Methods Thirty-six wildtype(C57BL/6)mice were selected to establish the orthodontic tooth movement model.The loading group was applied by 10-15 grams force between the first molar and incisor teeth in the left side of maxillary,while the control group was applied in the right side.9 mice were killed on the 1st day,the 7th day and the 14th day respectively.3 of them were made into paraffin tissue sections to observe morphology with immunohistochemical staining;3 of them were made into hard tissue sections(without decalcification)to show calcein bone deposition with Masson staining;3 of them were analyzed the express level of Sema3A gene with Western blotting.Results (1)Tooth movement was observed in all the experimental rats.It showed that the modeling establishment was successful.(2)In the control group,the periodontal tissue configuration was normal,there was no bone absorption or bone formation and other changes,and osteoblast is extremely rare.On the first day in the loading group,there was no obvious change in the periodontal tissue.On the 7th day,fiber membrane disorder was apparent.The periodontal ligament narrowing and a large number of osteoblast were visible.But on the 14th day,the morphology returned to normal with less osteoblast distribution.(3)Immunohistochemical staining showed the Sema3A expression was consistent negative in control group.In loading group,Sema3A expression was weak positive on the 1st day,then it rose to a strong positive expression on the 7th day,but it returned to weak positive on the 14th day.(4)Western blotting detection showed that there were Sema3A protein prints in two groups.Semi-quantitative analysis showed that there was no Sema3A protein expression fluctuation in control group,which meant in low level expression.In loading group,Sema3A protein expression was

关 键 词：正畸牙移动 牙周组织改建 骨改建 SEMA3A 小鼠 

分 类 号：R783.5[医药卫生—口腔医学]