白藜芦醇在人牙髓干细胞成牙本质向分化中的调控机制研究

doi:10.19439/j.sjos.2023.02.004

上海口腔医学 ›› 2023, Vol. 32 ›› Issue (2): 132-136.doi: 10.19439/j.sjos.2023.02.004

白藜芦醇在人牙髓干细胞成牙本质向分化中的调控机制研究

李宗谕, 李晓琳, 尉鹏功, 曲柳, 仇丽鸿, 于雅琼

中国医科大学口腔医学院·附属口腔医院牙体牙髓病科,辽宁省口腔疾病重点实验室,辽宁沈阳 110002

收稿日期:2022-04-16 修回日期:2022-06-23 出版日期:2023-04-25 发布日期:2023-06-13
通讯作者: 于雅琼,E-mail: yuyaqiong1987@126.com
作者简介:李宗谕（1998-）,女,在读硕士研究生,E-mail：lzy3515422186@163.com
基金资助:
辽宁省高等学校基本科研项目（青年项目）（LQNK201721）; 辽宁省自然科学基金指导计划（20180550532）

Regulation mechanism of resveratrol on odontogenic differentiation of human dental pulp stem cells

LI Zong-yu, LI Xiao-lin, WEI Peng-gong, QU Liu, QIU Li-hong, YU Ya-qiong

Department of Endodontics, School and Hospital of Stomatology, China Medical University; Liaoning Provincial Key Laboratory of Oral Diseases. Shenyang 110002, Liaoning Province, China

Received:2022-04-16 Revised:2022-06-23 Online:2023-04-25 Published:2023-06-13

摘要/Abstract

摘要： 目的: 探讨白藜芦醇是否通过上调沉默信息调节因子1（silent information regulator 1,SIRT1）的表达并激活β-catenin信号通路,从而促进人牙髓干细胞（DPSCs）成牙本质向分化。方法: 不同浓度白藜芦醇（0、10、15、20、50 μmol/L）作用于DPSCs 7天和14天后,利用CCK-8法检测细胞增殖活性。在15 μmol/L白藜芦醇作用下,成牙本质向分化诱导培养DPSCs 7 天后,检测碱性磷酸酶（ALP）活性;采用实时定量反转录PCR（qRT-PCR）检测Runt 相关转录因子2（Runx2）、牙本质涎磷蛋白（DSPP）和牙本质基质蛋白1（DMP-1）的mRNA表达情况;采用蛋白质免疫印迹法(Western blot)检测DPSCs分化诱导不同时间（0、3、5、7、14 天）后SIRT1的蛋白表达情况。15 μmol/L白藜芦醇作用下分化诱导培养DPSCs 7 天后,检测SIRT1和活化β连环蛋白（β-catenin）表达水平。采用GraphPad Prism 9软件包对数据进行统计学分析。结果: 15 μmol/L白藜芦醇对DPSCs 7天和14天的增殖活性无明显影响; 15 μmol/L白藜芦醇促进DPSCs的成牙本质向分化,上调Runx2、DSPP和DMP-1的mRNA表达。DPSCs分化诱导7 天时,SIRT1蛋白表达量最大;15 μmol/L白藜芦醇作用下分化诱导DPSCs 7 天时,SIRT1及活化β-catenin蛋白表达显著增加。结论: 白藜芦醇通过上调 SIRT1蛋白表达并激活β-catenin信号通路,促进人DPSCs成牙本质向分化。

关键词: 白藜芦醇, 牙髓干细胞, 成牙本质向分化, 沉默信息调节因子1, β连环蛋白

Abstract: PURPOSE: To investigate whether resveratrol promotes odontogenic differentiation of human dental pulp stem cells(DPSCs) by up-regulating the expression of silent information regulator 1 (SIRT1) and activating β-catenin signaling pathway. METHODS: Different concentrations of resveratrol(0, 10, 15, 20 and 50 μmol/L) were used to treat DPSCs for 7 days and 14 days, and cell proliferative activity was detected by CCK-8. After odontogenic differentiation induced by 15 μmol/L resveratrol for 7 days, alkaline phosphatase(ALP) staining was performed and real-time quantitative reverse transcription PCR(qRT-PCR) was used to detect the mRNA expression of Runt-related transcription factor 2 (Runx2), dentin sialophosphoprotein(DSPP) and dentin matrix protein-1(DMP-1) in DPSCs. Western blot was used to detect the expression of SIRT1 in DPSCs on a specific day (0, 3rd, 5th, 7th and 14th) after differentiation induction. Western blot was also used to detect the expression of SIRT1 and activated β-catenin during odontogenic differentiation of DPSCs treated by 15 μmol/L resveratrol for 7 days. The experimental data was analyzed with GraphPad Prism 9 software package. RESULTS: 15 μmol/L resveratrol had no significant effect on proliferation of DPSCs on the 7th and 14th day; 15 μmol/L resveratrol promoted odontogenic differentiation of DPSCs and up-regulated mRNA expression of RUNX2, DSPP, and DMP-1 in DPSCs; the expression of SIRT1 was the highest on the 7th day during odontogenic differentiation induction. Resveratrol resulted in the increasing protein expressions of SIRT1 and activated β-catenin when DPSCs was induced to odontogenic differentiation for 7 days. CONCLUSIONS: Resveratrol promotes odontogenic differentiation of human DPSCs by up-regulating the expression of SIRT1 protein and activating β-catenin signaling pathway.

Key words: Resveratrol, Dental pulp stem cells, Odontogenic differentiation, Silent information regulator 1, β-catenin

中图分类号:

R781.3

李宗谕, 李晓琳, 尉鹏功, 曲柳, 仇丽鸿, 于雅琼. 白藜芦醇在人牙髓干细胞成牙本质向分化中的调控机制研究[J]. 上海口腔医学, 2023, 32(2): 132-136.

LI Zong-yu, LI Xiao-lin, WEI Peng-gong, QU Liu, QIU Li-hong, YU Ya-qiong. Regulation mechanism of resveratrol on odontogenic differentiation of human dental pulp stem cells[J]. Shanghai Journal of Stomatology, 2023, 32(2): 132-136.

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白藜芦醇在人牙髓干细胞成牙本质向分化中的调控机制研究

Regulation mechanism of resveratrol on odontogenic differentiation of human dental pulp stem cells

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