【作者】 郭青玉； 肖刚； 李睿； 吴军正；

【Author】 GUO Qingyu;XIAO Gang, , LI Rui, WU Jun-zheng.(Stomatological Hospital, Xi’an Jiaotong University, Xi’an ;Department of Oral Biology,the Fouth Military University. Xi’an 710004, Shanxi Province, China)

【机构】 西安交通大学口腔医院口腔内科教研室陕西西安； 第四军医大学口腔医学院口腔生物学教研室；

【摘要】 目的:本研究探索硫化修饰的、靶向gtfB mRNA的反义脱氧寡核苷酸对变形链球菌的作用。方法:采用阳离子聚合体 So-fastTM为载体,硫代修饰的寡核苷酸转化变形链球菌,RT-PCR检测gtfB mRNA转录水平的变化,SDS-PAGE检测GtfB表达的变化,苯酚-硫酸法检测水不溶性葡聚糖合成的变化,电镜观察生物膜结构的变化。利用生长曲线,观察转化试剂和寡核苷酸对细菌增殖能力的影响。结果:浓度为10μmol/L的反义寡核苷酸处理变形链球菌后,可以抑制变形链球菌gtfB mRNA的转录, GtfB的表达和水不溶性葡聚糖的合成。而对于变形链球菌形成的生物膜,则可使其结构发生改变,包括基质的减少,更加分散的分布,粘附细胞层数的减少。并且反义寡核苷酸处理后的细菌生长速率并未受到影响。结论:反义技术有望成为新型的抗龋试剂。更多还原

【Abstract】 OBJECTIVE: To examine the effects of phosphorothioate-modified antisense oligodeoxyribonucleotides (PS-ODN) targeted to mRNA transcribed from gtfB gene in Streptococcus mutans. METHODS: Antisense PS-ODN were introduced into the S. mutans through So-fastTM package. Which is a transfection reagent composed of cationic polymer. RT-PCR was used to characterize the transcription of gtfB mRNA of S.mutans. Streptococcus mutans cell-associated proteins were resolved by SDS-PAGE.The total amount of insoluble glucans were measured by the phenol-sulfuric acid method. The architecture of S.mutans biofilm was observed by a scanning electron microscope.For PS-ODN and So-fastTM reagent, we observed its toxicity by means of growth curve. RESULT: Treatment of S. mutans with 10 μmol/L antisense PS-ODNs inhibited gtfB mRNA transcription. GtfB expression and water-insoluble glucan synthesis. The architecture of biofilms formed by antisense PS-ODNs-treated S.mutans showed reduced biomass, more dispersed distribution with enlarged interspaces and fewer layers of attached cells. PS-ODN treatment had no effect on the growth of S.mutans. CONCLUSION: Our results indicated that it might be feasible to use antisense PS-ODN as a novel agent in caries prevention.更多还原