华西口腔医学杂志

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尼古丁抑制成牙本质细胞增殖及作用机制的研究

吴礼安 文玲英 杨富生 王小竞   

  1. 第四军医大学口腔医院儿童口腔科, 陕西西安710032
  • 收稿日期:2008-04-25 修回日期:2008-04-25 出版日期:2008-04-20 发布日期:2008-04-20
  • 通讯作者: 文玲英,Tel: 029- 84776087
  • 作者简介:吴礼安(1975-), 男,安徽人,讲师,博士
  • 基金资助:

    第四军医大学学术新人基金资助项目(2006)

Effects and molecular mechanism of nicotine on odontoblasts

WU Li- an, WEN Ling- ying, YANG Fu- sheng, WANG Xiao- jing   

  1. Dept. of Pediatric Dentistry, College of Stomatology, The Fourth Military Medical University, Xi′an 710032, China
  • Received:2008-04-25 Revised:2008-04-25 Online:2008-04-20 Published:2008-04-20
  • Contact: WEN Ling- ying,Tel: 029- 84776087

摘要:

目的观察尼古丁对成牙本质细胞增殖的抑制作用,并检测其对成牙本质细胞中Ca2+浓度的影响,以探讨尼古丁抑制成牙本质细胞的分子机制。方法体外培养成牙本质细胞系MDPC- 23细胞, 按每毫升2×104个接种,随机分为实验组和对照组,对照组不加任何刺激,实验组施加质量浓度为100 μg/mL尼古丁,并于8 h后加入浓度为10 μmol/L BrdU进行细胞周期标记,刺激24 h后固定细胞,行免疫荧光抗BrdU染色,碘化丙锭(PI)复染胞核,荧光显微镜下计数细胞总数与BrdU阳性细胞数,计算S期阳性细胞率并进行统计学分析。培养成牙本质细胞于特制培养皿中,施加质量浓度为100 μg/mL尼古丁刺激,激光共聚焦显微镜下检测成牙本质细胞中Ca2+浓度的动态变化。结果实验组、对照组S期阳性细胞率分别为36.3%、48.2%,实验组显著低于对照组。尼古丁刺激后,成牙本质细胞中Ca2+浓度迅速升高,在较高水平维持一段时间后缓慢下降。结论尼古丁可抑制成牙本质细胞增殖,这种作用同尼古丁升高成牙本质细胞中Ca2+浓度有关。

关键词: 尼古丁, 成牙本质细胞, 增殖

Abstract:

Objective To observe the effects of nicotine on the proliferation of odontoblasts and explore the possible mechanism. Methods Odontoblasts MDPC- 23 were cultured, inoculated and divided into two groups randomly. With no stimuli added for the control group, the experimental group was stimulated by 100 μg/mL nicotine. After 8 hours, 10 μmol/L BrdU was added to label cells at S stage in cell cycle. 24 hours later, odontoblasts were fixed and immunofluorescence staining was performed with specific mouse BrdU antibody. After counterstaining with propidium iodide, BrdU positive cells were arbitrarily scored microscopically by an independent estimation conducted three times, and the corresponding total cell number in the same vision were counted in both groups. BrdU positive cell rates were calculated and compared statistically. At the same time, odontoblasts MDPC- 23 were cultured and stimulated by 100 μg/mL nicotine, the dynamic Ca2 + concentration inside the cytoplasm were detected immediately by a confocal laser scanning microscope. Results The ratio of S stage cells in the experimental group was 36.3% significantly lower than that(48.2%) in the control group. After the addition of 100 μg/mL nicotine, the Ca2+ concentration inside the cytoplasm rose rapidly, sustained at a high level for a short time and then relapsed gradually. Conclusion Nicotine had inhibitory effects on the proliferation of odontoblasts MDPC- 23, which might be related to the increased Ca2 +concentration in the cytoplasm.

Key words: nicotine, odontoblast, proliferation