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

doi:10.19439/j.sjos.2020.06.004

上海口腔医学 ›› 2020, Vol. 29 ›› Issue (6): 580-585.doi: 10.19439/j.sjos.2020.06.004

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

李琴^1,2, 吕春华^2,3, 陈玲^2,4, 刘贤^1,2, 王华², 孙雯²

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

收稿日期:2019-03-17 修回日期:2019-05-15 出版日期:2020-12-25 发布日期:2021-01-08
通讯作者: 孙雯,E-mail：wensun@njmu.edu.cn
作者简介:李琴（1981-）,女,硕士,副主任医师,E-mail: lq.11@163.com
基金资助:
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）

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

LI Qin^1,2, LYU Chun-hua^2,3, CHEN Lin^2,4, LIU Xian^1,2, WANG Hua², SUN Wen²

1. Department of Operative Dentistry, Changzhou Stomatology Hospital. Changzhou 213003, Jiangsu Province;
2. Jiangsu Key Laboratory of Oral Diseases, Nanjing Medical University. Nanjing 210029, Jiangsu Province;
3. Department of Stomatology, Taizhou People's Hospital. Taizhou 225300, Jiangsu Province;
4. Department of Stomatology, Shanghai Tongren Hospital. Shanghai 200050, China

Received:2019-03-17 Revised:2019-05-15 Online:2020-12-25 Published:2021-01-08

摘要/Abstract

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。

关键词: 甲状旁腺激素相关蛋白, 牙槽骨, 骨原细胞, 重组蛋白

Abstract: PURPOSE: To synthetise mouse parathyroid hormone related protein 1-36 (PTHrP1-36) and to study its role in promoting alveolar bone formation. METHODS: pGEX-2TK/ 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-2TK/ 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 1mM 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.

Key words: Parathyroid hormone related protein, Alveolar bone, Osteoprogenitor, Synthetic protein

中图分类号:

R782.1

李琴, 吕春华, 陈玲, 刘贤, 王华, 孙雯. 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[J]. 上海口腔医学, 2020, 29(6): 580-585.

LI Qin, LYU Chun-hua, CHEN Lin, LIU Xian, WANG Hua, SUN Wen. Synthetic PTHrP1-36 and its role in promoting alveolar bone formation[J]. Shanghai Journal of Stomatology, 2020, 29(6): 580-585.

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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

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