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肾上腺髓质素对人牙髓干细胞增殖和分化的影响 |
朱强,许震宇,田刚,唐震,高建勇,邱小倩,蔚一博,汪大林* |
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(第二军医大学长海医院口腔科, 上海 200433 *通信作者) |
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摘要: |
目的 探讨肾上腺髓质素(adrenomedullin,ADM)对人牙髓干细胞(dental pulp stem cells,DPSCs)增殖、成牙本质分化能力的影响。方法 体外培养的人DPSCs传至第3代,培养液中加入不同浓度ADM(10-6、10-7及10-8 mol/L)处理细胞24 h,并设置空白对照组。流式细胞术检测细胞周期及细胞凋亡,定量PCR和蛋白质印迹法检测成牙本质分化标记物碱性磷酸酶(alkaline phosphatase,ALP)的mRNA及蛋白表达。结果 细胞周期检测结果显示,3种浓度ADM干预DPSCs后G2/S/M期细胞比例均较空白对照组上升(P<0.05),但各干预组间差异无统计学意义; 细胞凋亡检测结果显示,3种浓度ADM干预组的Annexin Ⅴ+/PI-标示细胞比例均较空白对照组下降(P<0.05),但各干预组间差异无统计学意义; 定量PCR及蛋白质印迹检测结果显示,DPSCs中ALP mRNA及蛋白表达较空白对照组上升(P<0.01),但各干预组间差异无统计学意义。结论 ADM具有促进DPSCs的增殖、抑制凋亡及促其向成牙本质分化的作用。 |
关键词: 牙髓干细胞 肾上腺髓质素 细胞增殖 细胞分化 |
DOI:10.3724/SP.J.1008.2014.00515 |
投稿时间:2014-02-06修订日期:2014-02-27 |
基金项目: |
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Effects of adrenomedullin on proliferation and differentiation of dental pulp stem cells |
ZHU Qiang,XU Zhen-yu,TIAN Gang,TANG Zhen,GAO Jian-yong,QIU Xiao-qian,WEI Yi-bo,WANG Da-lin* |
(Department of Stomatology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China *Corresponding authors.) |
Abstract: |
Objective To explore the effects of adrenomedullin (ADM) on the proliferation and odontogenic differentiation potential of dental pulp stem cells (DPSCs). Methods The 3rd generation of cultured human DPSCs were treated with ADM of different concentrations (10-6 mol/L, 10-7 mol/L and 10-8 mol/L) for 24 h, and a blank control group was also set up. Cell apoptosis and cell cycle were measured by flow cytometry; the mRNA and protein expressions of alkaline phosphatase (ALP), an odontogenic differentiation marker, were analyzed by qPCR and Western blotting analysis, respectively. Results Cell cycle analysis showed that the proportion of G2/S/M phase cells in the three ADM treatment groups were significantly higher than that in the blank control group (P<0.05), but there were no significant differences between the three ADM groups. Cell apoptosis analysis showed that the proportion of cells labelled by Annexin Ⅴ+/PI- was significantly lower in the three ADM groups than that in the blank control group (P<0.05), but there were no signficant differences between the three ADM groups.The results of qPCR and Western blotting analysis showed that the ALP expression in DPSCs was increased significantly compared with that in the blank control group (P<0.01), but there were no significant differences between the three ADM groups. Conclusion ADM can promote the proliferation and inhibit the apoptosis of DPSCs, and it can also promote odontogenic differentiation of DPSCs. |
Key words: dental pulp stem cells adrenomedullin cell proliferation cell differentiation |