华西口腔医学杂志

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变异链球菌耐酸毒力因子质子移位膜ATP酶在龋病进展中的动态变化

高敬  黄文明   

  1. 重庆医科大学附属口腔医院牙体牙髓病科·口腔疾病与生物医学重庆市重点实验室·重庆市高校市级口腔生物医学工程重点实验室,重庆 401147
  • 收稿日期:2015-07-12 修回日期:2015-10-18 出版日期:2016-04-01 发布日期:2016-04-01
  • 通讯作者: 黄文明,讲师,硕士,E-mail:jxhwm@163.com
  • 作者简介:高敬,硕士,E-mail:326188065@qq.com
  • 基金资助:

    国家自然科学基金(31371473);重庆市卫生局医学科研计划(2012-2-129);重庆市卫生局医学科研计划重点项目(2011-1-062);重庆市渝北区科技项目(2011)

Dynamic changes of aciduric virulence factor membrane-bound proton-translocating ATPase of Streptococcus mutans in the development of dental caries

Gao Jing, Huang Wenming.   

  1. Dept. of Conservative Dentistry and Endodontics, Sto- matological Hospital of Chongqing Medical University; Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences; Chongqing Municipal Key Laboratory of Oral Biomedical Engineering Higher Education, Chongqing 401147, China
  • Received:2015-07-12 Revised:2015-10-18 Online:2016-04-01 Published:2016-04-01
  • Contact: Huang Wenming, E- mail: jxhwm@163.com.
  • Supported by:

    Natural Science Foundation of China (31371473); Medical Scientific Research Plan of Chongqing Municipal Health Bureau (2012-2-129); Key Program for Medical Scientific Research Plan of Chongqing Municipal Health Bureau (2011-1-062); Scientific and Technical Program of Chongqing Yubei District (2011).

摘要:

目的  研究变异链球菌耐酸毒力因子质子移位膜ATP酶(F-ATPase)在不同pH环境和龋病发生发展过程中的表达,评价F-ATPase在龋病进展中的动态变化。方法  将变异链球菌菌悬液在不同pH(pH4.0~7.0)和不同葡萄糖浓度(含5%和不含葡萄糖)的BHI液体培养基中培养,检测F-ATPase基因的表达水平。将雄性Wistar大鼠随机分成致龋组和对照组,其中致龋组喂养致龋饲料及5%葡萄糖水,对照组喂养普通饲料。每2周采集菌斑样本,检测F-ATPase基因的表达水平。第11周时取大鼠上下颌骨标本,对磨牙进行龋损评定。结果 1)5%葡萄糖浓度下变异链球菌F-ATPase基因的表达高于不含葡萄糖(P<0.05),在pH5.0时F-ATPase基因的表达最高,pH4.0时表达最低(P<0.05)。2)成功构建了龋病动物模型,在龋病发生发展过程中,致龋组和对照组F-ATPase的表达逐渐增强,致龋组表达高于对照组(P<0.05)。结论  耐酸毒力因子F-ATPase的表达变化与龋病的发生发展密切相关。

关键词: 龋病, 牙菌斑, 质子移位膜ATP酶, pH

Abstract:

Objective  To observe the dynamic changes of membrane-bound proton-translocating ATPase (F-ATPase) in the development of dental caries, the expression of Streptococcus mutans F-ATPase under different pH concentrations and during the development of dental caries is analyzed. Methods  Streptococcus mutans cultured under different pH (pH4.0-7.0) concentrations and containing 5% glucose and no glucose containing BHI were collected. RNA was extracted. Subsequently, F-ATPase gene was detected using real-time polymerase chain reaction. Male Wistar rats were divided randomly into caries group and control group. The rats in the caries group were fed caries feed and 5% glucose water, whereas those of control group were fed normal feed. Total RNA was extracted from plaque samples, which were collected from rats’ oral cavity every two weeks. F-ATPase gene was detected by real-time PCR. In the 11th week, the upper and lower jaw bone specimens of rats were taken, and molar caries damage assessed. Results  The expression of F-ATPase in the caries group was higher than that in the control group (P<0.05). In addition, the gene was expressed highest in pH5.0 and the lowest in pH4.0 (P<0.05). 2) The expression of F-ATPase progressively increased during the caries development in both groups; expression in the caries group was higher than that in control group (P<0.05). Conclusion  Acid-resisting viru-lence factor F-ATPase is related closely with the incidence and development of dental caries.

Key words: dental caries, dental plaque, membrane-bound proton-translocating ATPase, pH