体外观察反义脱氧寡核苷酸对变形链球菌蔗糖依赖性粘附的影响

The effect of gtfB specific antisense oligodeoxyribonucleotides on S.mutans sucrose-dependent adherence to saliva coated hydroxyapatite in vitro

目的：评估gtfB特异反义脱氧寡核苷酸对变形链球菌蔗糖依赖性粘附能力的影响。方法：合成针对gtfB709—726bp及3479—3497bp的反义脱氧寡核苷酸并进行硫化。将其分别自然转入变形链球菌细胞内，观察各组变形链球菌对唾液包被羟磷灰石的蔗糖依赖性粘附，结晶紫染色，无水乙醇脱色，酶标仪于620nm处读取吸光度值。结果：培养基中加入反义脱氧寡核苷酸的变形链球菌蔗糖依赖性粘附明显低于对照组（P〈0．01），PS-ODN1和PS-ODN2分别可减少31．4％和41．8％细菌粘附。结论：反义脱氧寡核苷酸可以抑制变形链球菌蔗糖依赖性粘附，有可能将其开发成龋病预防及病因研究中的新型试剂。

Objective ：To evaluate the effect of gtfB specific antisense phosphorothioate-modified oligodeoxyribonucleotides（PS-ODNs） on S. mutans sucrose-dependent adherence. Methods. Antisense oligodeoxyribonucleotide targeted to gtfB sequence 709 - 726 bp（ PS-ODN1 ） and 3 479 - 3 497 bp（ PS-ODN2 ） were synthesized. Natural genetic transformation of S. mutans with PS-ODNI and PS-ODN2 was respectively performed. Adhesion of S. mutans to saliva coated hydroxyapatite was examined by crystal violet staining and destain with absolute ethanol. The absorbance at 620 nm was measured by plate reader（ the absorbance value derived from the wells without sucrose was used as background and was subtracted）. Results：The adhesion ability of the strains treated with antisense PS-ODN was significantly lower than that of the control（P 〈 0.01 ）, PS-ODN1 and PS-ODN2 decreased the adhesion by 31.4% and 41.8% respectively. Conclusion：gtfB specific antisense ODNs can inhibit S. mutans sucrose-dependent adherence.