重组甲状旁腺激素相关蛋白1-36及其在促进牙槽骨骨形成中的作用  

Synthetic PTHrP1-36 and its role in promoting alveolar bone formation

作　　者：李琴[1,2] 吕春华[2,3] 陈玲[2,4] 刘贤[1,2] 王华[2] 孙雯 LI Qin;LYU Chun-hua;CHEN Lin;LIU Xian;WANG Hua;SUN Wen(Depariment of Operative Dentistry,Changzhou Stomatology Hospital,Changzhou 213003,Jiangsu Prowince;Jiangasu Key Laboratory of Oral Diseases,Nanjing Medical University,Nanjing 210029,Jiangsu Prowince;Department of Stomatology,Taizhou People's Hospital,Taizhou 225300,Jiangsu Prowince;Department of Stomatology,Shanghai Tongren Hospital,Shanghai 200050,China)

机构地区：[1]常州市口腔医院口腔内科,江苏常州213003 [2]南京医科大学口腔疾病研究江苏省重点实验室,江苏南京210029 [3]泰州市人民医院口腔科,江苏泰州225300 [4]上海同仁医院口腔科,上海200050

出　　处：《上海口腔医学》2020年第6期580-585,共6页Shanghai Journal of Stomatology

基　　金：江苏省口腔疾病研究重点实验室开放课题(JSKLOD-KF-1706);江苏省口腔疾病研究重点实验室开放课题(JSKLOD-KF-1707);高等学校博士学科点专项科研基金-新教师类(20123234120007)。

摘　　要：目的:构建重组甲状旁腺激素相关蛋白(parathyroid hormone related protein,PTHrP)1-36并观察其在促进牙槽骨骨形成中的作用。方法:构建pGEX-2TK/PTHrP1-36原核表达载体,表达重组小鼠PTHrP1-36蛋白。通过处理小鼠原代骨髓间充质细胞和卵巢切除致颌骨疏松的小鼠,体外和体内观察重组PTHrP1-36在促进牙槽骨骨形成中的作用。采用SPSS 10.0软件包对数据进行统计学分析。结果:实验结果显示,pGEX-2TK/PTHrP1-36原核表达载体构建成功,IPTG可诱导新生蛋白表达。体外处理小鼠骨髓细胞,发现重组PTHrP1-36蛋白可以增加总成纤维细胞克隆形成单位(CFU-f)、碱性磷酸酶阳性的CFU-f和钙化结节的数目。在卵巢切除致颌骨疏松的小鼠体内注射重组PTHrP1-36蛋白,发现小鼠牙槽骨骨密度增加。H-E染色以及骨胶原染色显示,牙槽骨骨量明显增加。结论:重组PTHrP1-36可促进骨髓间充质细胞的成骨分化,促进牙槽骨骨密度和骨量的增加,提示PTHrP1-36在促进牙槽骨形成中具有积极作用。PURPOSE:To synthetise mouse parathyroid hormone related protein 1-36(PTHrP1-36)and to study its role in promoting alveolar bone formation.METHODS:pGEX-2 TK/PTHrP1-36 was constructed and PTHrP1-36 was expressed.The role of PTHrP1-36 in promoting alveolar bone formation was observed by treating bone marrow cells in vitro and ovariectomy-induced maxillofacial osteoporosis in vivo.Statistical analysis was performed using SPSS 10.0 software package.RESULTS:The results of PCR analysis showed that pGEX-2 TK/PTHrP1-36 plasmid was correct templates for expressing PTHrP1-36.The molecular weights obtained by SDS-PAGE were the same as the theoretical value after IPTG induction.Scale up production was carried out by 3-hour-induction with 1 mM IPTG which was the optimal induction factors.Bone marrow cell culture revealed that synthetic PTHrP1-36 increased the number of total fibroblastic colony forming units(CFU-f),ALP positive CFU-f and calcific nodules.After PTHrP1-36 was subcutaneously administered to ovariectomy mice,density and volume of alveolar bone were improved,as showed by X-ray and histochemistry staining of collagen-l and H-E staining.CONCLUSIONS:Synthetic PTHrP1-36 can promote bone marrow cell osteogenic differentiation and increase alveolar bone formation.

关 键 词：甲状旁腺激素相关蛋白 牙槽骨 骨原细胞 重组蛋白 

分 类 号：R782.1[医药卫生—口腔医学]