上海口腔医学

上海口腔医学 ›› 2020, Vol. 29 ›› Issue (3): 225-230.doi: 10.19439/j.sjos.2020.03.001

• 论著 • 上一篇    下一篇

艾塞那肽对人牙周膜干细胞增殖、迁移和成骨分化的影响

梁乾宇1,2, 杜令倩3, 张睿1,2, 丁田1,2, 葛少华1,2   

  1. 1.山东大学口腔医学院 山东省口腔组织再生重点实验室, 山东 济南 250012;
    2.山东大学口腔医学院 牙周病科,山东 济南 250012;
    3.山东大学第二医院 口腔科,山东 济南 250033
  • 收稿日期:2019-01-07 修回日期:2019-04-14 出版日期:2020-06-25 发布日期:2020-07-29
  • 通讯作者: 葛少华,E-mail: shaohuage@sdu.edu.cn
  • 作者简介:梁乾宇（1993-）,男,硕士研究生,E-mail: 313340630@qq.com
  • 基金资助:
    国家自然科学基金（81670993,81873716）; 山东省重点研发计划（2018GSF118065）; 山东大学交叉学科培育项目（2018JC005）

Effects of Exenatide-4 on proliferation, migration and osteogenic differentiation of human periodontal ligament stem cells

LIANG Qian-yu1,2, DU Ling-qian3, ZHANG Rui1,2, DING Tian1,2, GE Shao-hua1,2   

  1. 1. Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of Stomatology, Shandong University. Jinan 250012;
    2. Department of Periodontology, School of Stomatology, Shandong University. Jinan 250012;
    3. Department of Stomatology, The Second Hospital of Shandong University. Jinan 250033, Shandong Province, China
  • Received:2019-01-07 Revised:2019-04-14 Online:2020-06-25 Published:2020-07-29

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摘要： 目的 探讨艾塞那肽（exendin-4, EX-4）对人牙周膜干细胞（periodontal ligament stem cells, PDLSCs)增殖、迁移和成骨分化的影响。方法 通过有限稀释法体外分离、培养PDLSCs。通过克隆形成实验,定向诱导成骨、成脂分化,鉴定其干细胞特性。通过免疫荧光染色检测PDLSCs表面EX-4受体GLP-1R（glucagon-like peptide-1 receptor）的表达。利用5、10、20或50 nmol/L EX-4体外刺激PDLSCs,通过CCK-8、Transwell实验和碱性磷酸酶（alkaline phosphatase, ALP）活性测试,研究不同浓度EX-4对PDLSCs增殖、迁移和成骨分化的影响。应用实时定量PCR检测成骨相关基因ALP、runt相关转录因子2（runt related transcription factor 2, Runx2）和骨钙素（osteocalcin, OCN）的表达。采用Graphpad Prims 6.0软件包对数据进行统计学分析。结果 成功分离、培养PDLSCs。GLP-1R在PDLSCs表面阳性表达,不同浓度EX-4对PDLSCs的增殖能力无显著影响（P>0.05）,10 nmol/L EX-4可显著促进PDLSCs迁移、ALP活性及成骨相关基因的表达（P<0.05）。结论 10 nmol/L EX-4可促进PDLSCs迁移和成骨分化。

关键词: 牙周膜干细胞, 艾塞那肽, 成骨分化

Abstract: PURPOSE: To investigate the effects of exendin-4(EX-4) on proliferation, migration and osteogenic differentiation of human periodontal ligament stem cells(PDLSCs). METHODS: PDLSCs were isolated and cultured using limited dilution method in vitro. Colony formation assay, osteogenic and adipogenic differentiation were applied to identify the stem cells. Immunofluorescence staining was used to detect the expression of EX-4 receptor glucagon-like peptide-1 receptor (GLP-1R) on the surface of PDLSCs. PDLSCs were stimulated with 5, 10, 20 or 50 nmol/L EX-4 in vitro. CCK-8, Transwell assay and alkaline phosphatase(ALP) activity assay were used to determine the effects of EX-4 on PDLSCs proliferation, migration and osteogenic differentiation. Quantitative real-time polymerase chain reaction was used to determine the expression of osteogenic related genes ALP, runt-related transcription factor 2(Runx2) and osteocalcin (OCN). The data were analyzed by Graphpad Prims 6.0 software package. RESULTS: PDLSCs were successfully isolated and cultivated. GLP-1R positively expressed on the surface of PDLSCs. EX-4 exerted no significant effect on PDLSCs proliferation(P>0.05). EX-4 significantly promoted migration, ALP activity and osteogenic related genes expression of PDLSCs (P<0.05). CONCLUSIONS: 10 nmol/L EX-4 could promote migration and osteogenic differentiation of PDLSCs.

Key words: Periodontal ligament stem cells, Exendin-4, Osteogenic differentiation

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