黄连素促人牙髓干细胞成骨/成牙本质细胞向分化

doi:10.13701/j.cnki.kqyxyj.2020.07.008

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (7): 639-643.DOI: 10.13701/j.cnki.kqyxyj.2020.07.008

黄连素促人牙髓干细胞成骨/成牙本质细胞向分化

马兰¹, 禹怡君¹, 刘含笑¹, 刘超², 苗雷英^1*

1.南京大学医学院附属口腔医院牙体牙髓科江苏南京 210008;
2.南京大学医学院附属口腔医院正畸科江苏南京 210008

收稿日期:2019-11-15 出版日期:2020-07-28 发布日期:2020-07-24
通讯作者: 苗雷英,E-mail: miaoleiying80@163.com
作者简介:马兰(1994~),女,浙江绍兴人,硕士在读,主要从事牙髓生物学相关研究。
基金资助:
国家自然科学基金面上项目(编号:81570952);科教强卫工程江苏省青年医学人才;(编号:QNRC201612);南京市医学科技发展项目(编号:ykk18126)

Berberine Stimulates Osteogenesis/Odontogenic Differentiation of Human Dental Pulp Stem Cells

MA Lan¹, YU Yijun¹, LIU Hanxiao¹, LIU Chao², MIAO Leiying^1*

1. Department of Endodontics, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing 210008, China;
2. Department of Orthodontics, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing 210008, China

Received:2019-11-15 Online:2020-07-28 Published:2020-07-24

摘要/Abstract

摘要： 目的: 研究黄连素对人牙髓干细胞(hDPSCs)增殖及成骨/成牙本质细胞向分化的影响,为促进年轻恒牙牙根继续发育提供新思路。方法: 分离培养hDPSCs,流式细胞术鉴定细胞表型,诱导染色评估多向分化潜能。用不同浓度黄连素(0、1、3、10、30 μmol/L)刺激hDPSCs,通过CCK-8法、碱性磷酸酶活性检测、q-PCR法检测各组细胞增殖和成骨/成牙本质细胞向分化能力。结果: 改良组织块酶消化法成功分离hDPSCs并对其鉴定。CCK-8结果显示培养7 d内,0~10 μmol/L浓度的黄连素对hDPSCs增殖无影响,30 μmol/L组在第7天时抑制细胞增殖。黄连素浓度为1、3 μmol/L时细胞碱性磷酸酶活性显著升高。q-PCR结果也显示黄连素能上调成骨和成牙本质相关基因mRNA表达,且促进效果在浓度为1和3 μmol/L时最显著。结论: 黄连素在1~3 μmol/L浓度范围内能促进hDPSCs成骨/成牙本质细胞向分化。

关键词: 黄连素, 人牙髓干细胞, 成骨分化, 成牙本质分化

Abstract: Objective: To investigate the effect of berberine on proliferation and osteogensis/odontogenic differentiation of human dental pulp stem cells. Methods: Human dental pulp stem cells (hDPSCs) were isolated and cultured. The phenotype was identified by flow cytometry. hDPSCs were treated with different concentrations of berberine (0, 1, 3, 10, and 30 μmol/L). Cell proliferation and osteogenesis/odontogenic differentiation were detected by CCK-8, ALP activity assay, and q-PCR. Results: hDPSCs were successfully isolated and identified. CCK-8 results showed that within 7 days of culture, berberine at 0-10 μmol/L concentration had no effect on hDPSCs proliferation. However, 30 μmol/L group inhibited cell proliferation at day 7. ALP activity assay detected that berberine significantly promoted the activity of alkaline phosphatase at the concentration of 1 and 3μmol/L. q-PCR also demonstrated that berberine could up-regulate the mRNA expression of genes related to osteogenesis and odontogenic, and the promotion effect was most significant when the concentration was 1 and 3 μmol/L. Conclusion: Berberine can promote osteogenesis/odontogenic differentiation of hDPSCs at 1-3 μmol/L.

Key words: berberine, human dental pulp stem cells, osteogenesis differentiation, odontogenic differentiation

马兰, 禹怡君, 刘含笑, 刘超, 苗雷英. 黄连素促人牙髓干细胞成骨/成牙本质细胞向分化[J]. 口腔医学研究, 2020, 36(7): 639-643.

MA Lan, YU Yijun, LIU Hanxiao, LIU Chao, MIAO Leiying. Berberine Stimulates Osteogenesis/Odontogenic Differentiation of Human Dental Pulp Stem Cells[J]. Journal of Oral Science Research, 2020, 36(7): 639-643.

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黄连素促人牙髓干细胞成骨/成牙本质细胞向分化

Berberine Stimulates Osteogenesis/Odontogenic Differentiation of Human Dental Pulp Stem Cells

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