Liang Qian-Yu, DU Ling-Qian, Zhang Rui, Ding Tian, Ge Shao-Hua
National Shandong Provincial Key Laboratory of Oral Tissue Regeneration,Department of Periodontology, School of Stomatology, Shandong University. Jinan 250012, China. E-mail:
Shanghai Kou Qiang Yi Xue. 2020 Jun;29(3):225-230.
To investigate the effects of exendin-4(EX-4) on proliferation, migration and osteogenic differentiation of human periodontal ligament stem cells(PDLSCs).
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.
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).
10 nmol/L EX-4 could promote migration and osteogenic differentiation of PDLSCs.
研究艾塞那肽-4（EX-4）对人牙周膜干细胞（PDLSCs）增殖、迁移和成骨分化的影响。
采用有限稀释法体外分离培养PDLSCs。通过集落形成试验、成骨和成脂分化鉴定干细胞。采用免疫荧光染色检测PDLSCs表面EX-4受体胰高血糖素样肽-1受体（GLP-1R）的表达。体外分别用5、10、20或50 nmol/L EX-4刺激PDLSCs。采用CCK-8法、Transwell试验和碱性磷酸酶（ALP）活性测定法检测EX-4对PDLSCs增殖、迁移和成骨分化的影响。采用定量实时聚合酶链反应检测成骨相关基因ALP、 runt相关转录因子2（Runx2）和骨钙素（OCN）的表达。数据采用Graphpad Prims 6.0软件包进行分析。
成功分离培养PDLSCs。GLP-1R在PDLSCs表面呈阳性表达。EX-4对PDLSCs增殖无显著影响（P>0.05）。EX-4可显著促进PDLSCs的迁移、ALP活性及成骨相关基因的表达（P<0.05）。
10 nmol/L EX-4可促进PDLSCs的迁移和成骨分化。
