冠方牙本质中5种基质金属蛋白酶的分布及定量检测  

Localization and quantitative detection of matrix metalloproteinase in human coronal dentine

作　　者：王丹杨[1] 张凌[2] 李芳[2] 马克强[3] 陈吉华[2] 

机构地区：[1]西安医学院口腔医学系口腔修复学教研室,710021 [2]第四军医大学口腔医学院修复科军事口腔医学国家重点实验室,西安710032 [3]广州军区武汉总医院口腔科

出　　处：《中华口腔医学杂志》2014年第11期688-692,共5页Chinese Journal of Stomatology

基　　金：国家自然科学基金（81130078、81100772、51373198）

摘　　要：目的 比较5种基质金属蛋白酶（matrix metalloproteinase,MMP）在人冠方牙本质中的分布情况,以期为与胶原降解相关的各种牙体硬组织疾病研究提供科学基础.方法 将10颗磨牙制成牙本质组织片60片,分为5组,每组再分成2个亚组（每亚组6片）,分别采用化学染色法和荧光染色法进行免疫组化染色,光学和荧光显微镜观察.将27颗磨牙纵切成牙本质片,制备浅层和深层牙本质粉末,借助液相芯片技术分别检测MMP-1、2、3、8、9含量.结果 5种MMP在牙本质中均呈阳性表达,深层牙本质染色更深,釉质牙本质界内侧存在MMP-2、8、9高表达条带.牙本质浅层和深层MMP-1、2、3、8、9含量分别为（0.037±0.025）和（0.433±0.089）、（0.445±0.115）和（2.730±0.712）、（0.071±0.069）和（0.460±0.108）、（0.586±0.246）和（6.159±0.948）、（0.384±0.185）和（1.460±0.251） ng/mg.深层牙本质中每种MMP含量均显著高于浅层（P＜0.05）.结论 人冠方牙本质含MMP-1、2、3、8和9,其分布规律为从牙本质深层至浅层含量逐渐降低.Objective To compare the distribution and concentrations of matrix metalloproteinase （MMP）-1,2,3,8,9 in human coronal dentin.Methods The localization of five types of MMP was performed using immunohistochemistry.Molars were demineralized and sectioned into 5 μm thick specimens.All specimens were randomly divided into five groups according to the antibodies.Each group contained two subgroups （n=6）.Immunoreactivity of each subgroup was visualized with 3,3-diaminobenzidine solution or fluorescein isothiocyanate and observed under microscopy respectively.Molars were sectioned into slices.The slices were divided into two groups according to superficial or deep dentin and pulverized to fine powder.After dentin protein was extracted,the concentrations of MMP-1,2,3,8,9 were detected by using fluorescent microsphere immunoassay.Results Immunohistochemical staining revealed that MMP-1,2,3,8,9 were highly concentrated in the deep dentin.However,intense immunoreactivities of MMP-2,8,9 were identified in a 6-10 μm wide zone adjacent to the dentino-enamel junction.The content of MMP-1 in superficial layer and deep layer of dentin were （0.037 ± 0.025） and （0.433 ± 0.089） ng/mg.The content of MMP-2 in superficial layer and deep layer of dentin were （0.445 ±0.115） and （2.730±0.712） ng/mg.The content of MMP-3 in superficial layer and deep layer of dentin were （0.071 ±0.069） and （0.460±0.108） ng/mg.The content of MMP-8 in superficial layer and deep layer of dentin were （0.586±0.246） and （6.159±0.948） ng/mg.The content of MMP-9 in superficial layer and deep layer of dentin were （0.384±0.185） and （1.460± 0.251） ng/mg.The concentrations of all tested MMP were significantly higher in deep dentin than those in superficial dentin （P＜0.05）.Conclusions There are five types of MMP contained in human coronal dentin,and the distribution of MMP shows a decreasing trend from the deep dentin to the superficial dentin.

关 键 词：基质金属蛋白酶类 牙本质 免疫组织化学 

分 类 号：R783.3[医药卫生—口腔医学]