炎症微环境下骨髓间充质干细胞骨向分化中Periostin基因表达研究

doi:10.13701/j.cnki.kqyxyj.2020.12.015

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (12): 1148-1152.DOI: 10.13701/j.cnki.kqyxyj.2020.12.015

炎症微环境下骨髓间充质干细胞骨向分化中Periostin基因表达研究

卢永超^1,2, 王伟², 吕翱^1,2, 刘逸卿^2,3, 杜毅^2*

1.滨州医学院口腔医学院山东烟台 264003;
2.济南市口腔医院牙体牙髓科山东济南 250001;
3.大连医科大学口腔医学院辽宁大连 116044

收稿日期:2019-12-31 出版日期:2020-12-28 发布日期:2020-12-28
通讯作者: *杜毅,E-mail:dykqyy@163.com
作者简介:卢永超(1992~ ),男,菏泽人,硕士在读,研究方向:牙体牙髓牙周病学。
基金资助:
济南市卫健委科技计划项目(编号:2018-1-33)

Expression of Periostin Gene in Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells in Inflammatory Microenvironment

LU Yongchao^1,2, WANG Wei², LV Ao^1,2, LIU Yiqing^2,3, DU Yi^2*

1. School of Stomatology, Binzou Medical University, Yantai 264003, China;
2. Department of Oral Medicine, Jinan Stomatological Hosipital, Jinan 250001, China;
3. School of Stomatology, Dalian Medical University, Dalian 116044, China

Received:2019-12-31 Online:2020-12-28 Published:2020-12-28

摘要/Abstract

摘要： 目的:探讨炎症微环境对Wistar大鼠骨髓间充质干细胞(rBMSCs)成骨分化Periostin基因表达的影响。方法:采用全骨髓贴壁法分离和培养rBMSCs,取第3代rBMSCs成骨、成脂分化诱导;10 μg/L TNF-α作用于rBMSCs模拟炎症微环境,采用CCK-8法检测细胞增殖活性,qRT-PCR 检测 IL-1β及 TNF-α的基因表达。将第3代rBMSCs分成对照组(OM)、炎症组(OM+TNF-α)进行成骨诱导,茜素红染色及钙离子沉积量检测,在成骨诱导的第7、14天采用qRT-PCR检测细胞中Periostin和Runx2的基因表达水平,Western blot检测Periostin蛋白表达量。结果:全骨髓贴壁法成功分离培养rBMSCs;炎症组细胞TNF-α及IL-1β的基因表达水平分别是对照组的1.8倍(P<0.05)、2.1倍(P<0.01);炎症组较对照组成骨能力下降,相关成骨基因表达量降低;成骨诱导后炎症组的Periostin基因及蛋白表达水平较对照组明显降低(P<0.05)。结论:炎症微环境下诱导rBMSCs成骨分化Periostin基因表达水平下降,且rBMSCs的成骨能力降低。

关键词: 骨髓间充质干细胞, 炎症, 成骨分化, Periostin

Abstract: Objective: To investigate the expression level of Periostin gene in osteogenic differentiation of wistar rat bone marrow mesenchymal stem cells (rBMSCs) in inflammatory microenvironment. Methods: rBMSCs were isolated and cultured by whole bone marrow adherence method. The cell morphology was observed and the osteogenic differentiation of the third generation rBMSCs was induced. The cell inflammatory response was induced by 10ng/mL TNF-α. Then, the cell proliferation activity was detected by CCK-8 and the gene expression levels of IL-1β and TNF-α were measured by real-time quantitative PCR. The third generation of rBMSCs was divided into control group and inflammation group for osteogenic induction. The deposition amounts of calcium ions were measured after alizarin red staining. The expression levels of Periostin in the cells were detected by qRT-PCR and Western blot on the 14th day of osteogenic induction. Results: rBMSCs were successfully isolated and cultured by whole bone marrow adherence method. The gene expression levels of TNF-α and IL-1β in the inflammation group were 1.8 times (P<0.05) and 2.1 times (P<0.01) of those of control group, respectively. Compared with the control group, the inflammation group had lower bone formation ability and lower expression of related osteogenic genes. The expression levels of Periostin gene and protein in the inflammation group after osteoinduction were significantly lower than those in the control group (P<0.05). Conclusion: The expression level of Periostin gene in osteogenic differentiation of rBMSCs induced by inflammatory microenvironment decreased, and the osteogenic capacity of rBMSCs decreased.

Key words: bone marrow mesenchymal stem cells, inflammation, osteogenic differentiation, Periostin

卢永超, 王伟, 吕翱, 刘逸卿, 杜毅. 炎症微环境下骨髓间充质干细胞骨向分化中Periostin基因表达研究[J]. 口腔医学研究, 2020, 36(12): 1148-1152.

LU Yongchao, WANG Wei, LV Ao, LIU Yiqing, DU Yi. Expression of Periostin Gene in Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells in Inflammatory Microenvironment[J]. Journal of Oral Science Research, 2020, 36(12): 1148-1152.

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炎症微环境下骨髓间充质干细胞骨向分化中Periostin基因表达研究

Expression of Periostin Gene in Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells in Inflammatory Microenvironment

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