活化素Ⅰ型受体对小鼠下颌骨髁突软骨生长发育的影响及其机制  

Effect of activin A receptor typeⅠon growth and development of mandibular condylar cartilage in mice and its mechnism

作　　者：闫广兴 叶佳朋 王爽爽 刘苍维 周怡君 胡月[1] 郝新青[1] 杜奥博 史册[1] 孙宏晨[1,3] YAN Guangxing;YE Jiapeng;WANG Shuangshuang;LIU Cangwei;ZHOU Yijun;HU Yue;HAO Xinqing;DU Aobo;SHI Ce;SUN Hongchen(Department of Pathology, Stomatology Hospital, Jilin University, Changchun 130021, China;Department of Oral and Maxillofacial Surgery, Stomatology Hospital, Jilin University, Changchun 130021, China;Department of Pathology, Affiliated Stomatology Hospital, China Medical University, Shenyang110001, China;Department of Operative and Endodontics, Stomatology Hospital, Jilin University, Changchun 130021, China)

机构地区：[1]吉林大学口腔医院病理科,吉林长春130021 [2]吉林大学口腔医院颌面外科,吉林长春130021 [3]中国医科大学附属口腔医院口腔病理科,辽宁沈阳110001 [4]吉林大学口腔医院牙体牙髓科,吉林长春130021

出　　处：《吉林大学学报（医学版）》2020年第3期492-497,共6页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81920108012,81970903);吉林省财政厅科研项目资助课题(JCSZ2019378-6);吉林省卫生厅科研项目资助课题(2019Q013);中国博士后科学基金资助课题(2018T110258,2017M621219);吉林大学2018年研究生创新研究计划项目资助课题(101832018C077)。

摘　　要：目的:观察Ⅰ型骨形态发生蛋白(BMP)受体活化素Ⅰ型受体(ACVR1)对出生后小鼠下颌骨髁突软骨(MCC)细胞形态、增殖和分化的影响,为研究MCC相关性疾病的病因及治疗提供参考。方法:采用Cre-LoxP系统构建在C57BL/6J小鼠MCC细胞中条件性敲除ACVR1基因的动物模型。将基因型为Acvr1fx/fx;RS/RS和Acvr1+/-;Osterix(+)/(-)的雌性和雄性小鼠配对合笼,以子代Osterix-Cre(+);Acvr1fx/-;RS/+基因型小鼠为实验组,Osterix-Cre(+);Acvr1fx/+;RS/+小鼠为对照组。分别取新出生(n=3)、出生后21天(PN21)(n=4)和出生后42天(PN42)(n=5)雄性小鼠,X-gal染色检测MCC组织中Osterix-Cre表达,micro-CT法检测2组小鼠下颌骨髁突宽度和髁头长度,HE染色和甲苯胺蓝染色分析2组小鼠MCC细胞形态及各层软骨厚度,免疫组织化学(IHC)染色检测2组小鼠MCC组织中增殖细胞核抗原(PCNA)阳性细胞数和Ⅹ型胶原水平。结果:X-gal染色和IHC染色,ACVR1条件性基因缺失小鼠模型构建成功。在PN21时,与对照组比较,实验组小鼠下颌骨髁突宽度和髁头长度明显变短(P<0.05);2组小鼠MCC细胞形态表现和结构无明显差异。与对照组比较,实验组小鼠肥大软骨细胞层(Hy)和成软骨细胞层(Ch)这2层及Hy单层MCC组织中PCNA阳性细胞数明显增多(P<0.05或P<0.01)。在PN42时,与对照组比较,实验组小鼠部分下颌骨髁突软骨细胞形态异常,部分髁突肥大软骨细胞的排列紊乱,实验组小鼠髁突软骨前1/3区的Hy和中1/3区除Hy以外的其他层细胞厚度明显增大(P<0.05或P<0.01);与对照组比较,实验组小鼠MCC组织各层PCNA阳性细胞数和Ch中Ⅹ型胶原水平明显升高。结论:ACVR1通过抑制MCC细胞增殖和成软骨细胞向肥大软骨细胞的分化,从而影响MCC细胞形态及MCC组织结构。Objective:To observe the effect of typeⅠbone morphogenetic protein(BMP)receptor activin A receptor typeⅠ(ACVR1)on the morphology,proliferation and differentiation of the mandibular condylar cartilage(MCC)cells in the postnatal mice,and to provide the reference for the study on etiology and treatment of MCC-related disease.Methods:The C57BL/6J mouse model of conditional deletion of ACVR1 gene was constructed by using the Cre-LoxP system.The female and male mice with Acvr1fx/fx;RS/RS and Acvr1+/-;Osterix(+)/(-)genotypes were paired off with each other;the offspring Osterix-Cre(+);Acvr1fx/-;RS/+genotype mice were selected as experiment group,and the Osterix-Cre(+);Acvr1fx/+;RS/+mice were selected as control group.The newborn(n=3),postnatal day 21(PN21)(n=4)and PN42(n=5)male mice were selected.X-gal staining was used to detect the expressions of Osterix-Cre in MCC tissue of the mice in two groups,micro-CT was used to detect the condylar widths and condylar head lengths of mandible of the mice in two groups,HE and Toluidine blue staining were used to analyze the morphology of MCC cells and the thickness of caritilage in each layer of MCC tissue of the mice in two groups,immunohistochemical(IHC)staining was used to detect the number of proliferating cell nuclear antigen(PCNA)-positive cells and the level of typeⅩcollagen in MCC tissue of the mice in two groups.Results:The X-gal staining and IHC results showed that the mouse model of ACVR1 gene conditional deletion was successfully constructed.At PN21,compared with control group,the condylar width and the condylar head length of mandible of the mice in experiment group were significantly shortened(P<0.05);the morphology of the MCC cells of the mice in two groups had no significant difference.Compared with control group,the number of PCNA-positive cells in the MCC cells of hypertrophic chondrocyte zone(Hy)and chondroblastic zone(Ch)and single Hy of the mice in experiment group were significantly increased(P<0.05 or P<0.01).At PN42,compared with control group,the sh

关 键 词：活化素Ⅰ型受体 下颌骨髁突软骨 细胞增殖 细胞分化 成软骨细胞 肥大软骨细胞 

分 类 号：R782.1[医药卫生—口腔医学]