伊犁黑蜂蜂胶对口腔主要致龋细菌生物膜作用的实验研究

Effects of Yili dark bee propolis on oral cariogenic biofilm in vitro

目的研究伊犁黑蜂蜂胶对口腔常见致龋细菌生物膜的影响,探讨其防龋效果及可能的防龋机制。方法通过结晶紫染色法测定黑蜂蜂胶对口腔常见致龋菌(变异链球菌、表兄链球菌、血链球菌、黏性放线菌、内氏放线菌)的最小生物膜清除浓度(MBEC);培养测试细菌24 h单菌生物膜,加入MBEC及以下的3个浓度配置成初始p H值为7.0的含药培养基,厌氧培养24 h后测p H值,并计算p H变化值以检测不同浓度黑蜂蜂胶对测试菌单菌生物膜产酸能力的影响。蒽酮法测定MBEC及以下的3个浓度的黑蜂蜂胶对变异链球菌24 h单菌生物膜产生水不溶性胞外多糖的影响。结果黑蜂蜂胶对变异链球菌、表兄链球菌、血链球菌、黏性放线菌、内氏放线菌的MBEC分别是6.25、1.56、3.13、0.78、0.78 mg·m L-1;黑蜂蜂胶可使各测试菌单菌生物膜Δp H降低,蜂胶各组与对照组相比差异均有统计学意义(P<0.05);在MBEC浓度时,蜂胶可使变异链球菌合成水不溶性胞外多糖的能力降低。结论伊犁黑蜂蜂胶具有一定的防龋效果,其可能的防龋机制是通过有效清除口腔主要致龋细菌单菌生物膜,抑制测试菌株产酸、合成水不溶性胞外多糖的能力起作用的。

Objective To evaluate the effects of Yili dark bee propolis on the main cariogenic biofilm and mechanisms. Methods Susceptibilities to the ethanolic extract of propolis against Streptococcus mutans（S. mutans）, Streptococcus sobrinus（S. sobrinus）, Streptococcus sanguis（S. sanguis）, Actinomyces viscosus（A. viscosus）, and Actinomyces naeslundii（A. naeslundii） were analyzed by crystal violet stain method to determine the minimum biofilm eradication concentration（MBEC）. The biofilm was initially cultivated for 24 h. Subsequently, the propolis groups with different concentration MBEC and initial p H 7.0 were cultured for 24 h. Moreover, the p H value was measured to evaluate the acid-producing ability of the tested plaque biofilm. The effects of propolis on the insoluble extracellular polysaccharide synthesis of S. mutans biofilm were evaluated by anthrone method. Results The MBEC of Yili propolis on S. mutans, S. sobrinus, S. sanguis, A. viscosus, and A. naeslundii were 6.25, 1.56, 3.13, 0.78, and 0.78 mg·m L-1, respectively. Propolis could decrease the Δp H of the tested plaque biofilm, and the differences between the control and propolis groups were statistically significant（P0.05）. At MBEC, propolis could reduce the ability of S. mutans in synthesizing insoluble extracellular polysaccharides. Conclusion Yili propolis demonstrate remarkable eradicative effects on the cariogenic plaque biofilm, showing inhibition of the synthesis of biofilm-produced acids and insoluble extracellular polysaccharides.