重组人成纤维细胞生长因子21调节成牙骨质细胞矿化作用及其机制研究
Effect of recombinant human fibroblast growth factor 21 on the mineralization of cementoblasts and its related mechanism
作者：邬浩, 李莹, 王玉琢, 于继泽, 包幸福, 胡敏
Author：Wu Hao, Li Ying, Wang Yuzhuo, Yu Jize, Bao Xingfu, Hu Min
收稿日期：2022-09-21 年卷（期）页码：2023,41(2):140-140-148
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：成纤维细胞生长因子21,牙骨质再生,成牙骨质细胞,转化生长因子β/骨形态发生蛋白信号通路,
Key words：fibroblast growth factor 21,cementum regeneration,cementoblasts,transforming growth factor β/ bone morphogenetic protein signaling pathway,
基金项目：国家自然科学基金(81901036);吉林省科技发展计划项目(20200201358JC);吉林省财政厅医疗卫生人才项目(jcsz2020304-22);吉林大学研究生创新研究计划(101832020CX305)
中文摘要
目的 探讨重组人成纤维细胞生长因子21（rhFGF21）对成牙骨质细胞增殖和矿化的作用及其机制。 方法 采用苏木精-伊红染色、免疫组织化学染色和免疫荧光法检测成纤维细胞生长因子21（FGF21）在大鼠牙周组织和成牙骨质细胞OCCM-30中的表达和分布；采用CCK-8法检测经不同浓度rhFGF21处理OCCM-30的增殖情况；采用碱性磷酸酶染色和茜素红染色分别检测OCCM-30矿化诱导3、7 d后的矿化状态；通过实时定量聚合酶链反应（PCR）及蛋白免疫印迹法（Western blot）检测成骨相关基因Runx2和Osterix的转录及蛋白表达情况；通过PCR阵列分析评估OCCM-30内转化生长因子β（TGFβ）/骨形态发生蛋白（BMP）信号通路基因的表达变化。 结果 FGF21在大鼠牙周组织和OCCM-30中存在表达；rhFGF21对OCCM-30的增殖能力无明显影响，但50 ng/mL rhFGF21能促进OCCM-30矿化能力增强（P<0.001）；实时定量PCR结果显示Runx2和Osterix的转录水平在50 ng/mL rhFGF21矿化诱导3 d时上升，5 d时下降；Western blot结果显示，Runx2及Osterix的蛋白表达水平在矿化诱导过程中增强，rhFGF21上调细胞内Bmpr1b的表达。 结论 rhFGF21能够促进OCCM-30矿化，这与TGFβ/BMP信号通路的激活有关。
英文摘要
ObjectiveTo investigate the effect of recombinant human fibroblast growth factor 21 (rhFGF21) on the proliferation and mineralization of cementoblasts and its mechanism.MethodsHematoxylin eosin, immunohistochemical staining, and immunofluorescence were used to detect the expression and distribution of fibroblast growth factor 21 (FGF21) in rat periodontal tissues and cementoblasts (OCCM-30), separately. Cell Counting Kit-8 was used to detect the proliferation of OCCM-30 under treatment with rhFGF21. Alkaline phosphatase staining and Alizarin Red staining were used to detect the mineralization state of OCCM-30 after 3 and 7 days of mineralization induction. The transcription and protein expression of the osteogenic-related genes Runx2 and Osterix were detected by real-time quantitative polymerase chain reaction (PCR) and Western blot analysis. The expression levels of genes of transforming growth factor β (TGFβ)/bone morphogenetic protein (BMP) signaling pathway in OCCM-30 were detected through PCR array analysis.ResultsFGF21 was expressed in rat periodontal tissues and OCCM-30. Although rhFGF21 had no significant effect on the proliferation of OCCM-30, treatment with 50 ng/mL rhFGF21 could promote the mineralization of OCCM-30 cells after 7 days of mineralization induction. The transcriptional levels of Runx2 and Osterix increased significantly at 3 days of mineralization induction and decreased at 5 days of mineralization induction. Western blot analysis showed that the protein expression levels of Runx2 and Osterix increased during mineralization induction. rhFGF21 up-regulated Bmpr1b protein expression in cells.ConclusionrhFGF21 can promote the mineralization ability of OCCM-30. This effect is related to the activation of the TGFβ/BMP signaling pathway.
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