伊犁黑蜂蜂胶对主要致龋细菌及其生物膜作用的实验研究

发布时间：2018-06-07 09:15

本文选题：蜂胶 + 菌斑生物膜　；参考：《新疆医科大学》2014年硕士论文

【摘要】：目的：研究伊犁黑蜂蜂胶对口腔常见致龋菌及其生物膜毒力因子的作用,探讨其防龋效果及可能的防龋机制。方法：①通过液体稀释法测定伊犁黑蜂蜂胶对口腔常见致龋菌的最低抑菌浓度(MIC)及最低杀菌浓度(MBC)；②结晶紫染色法测定伊犁黑蜂蜂胶对口腔常见致龋菌的最小生物膜清除浓度(MBEC);③以MIC以下的4个浓度(包括MIC浓度)配置成初始pH为7.0的含药培养基,以空白液体培养基作为对照组,加入菌液厌氧培养后测其pH值,并计算pH变化值(△pH值)以检测不同浓度黑蜂蜂胶对浮游状态下测试菌产酸能力的影响；④葸酮法测定MIC以下4个浓度(包括MIC浓度)的黑蜂蜂胶对变形链球菌、粘性放线菌产生胞外多糖的影响,空白液体培养基作为对照组；⑤培养测试细菌24h单菌生物膜,加入MBEC以下的四个浓度(包括MBEC浓度)配置成初始pH值为7.0的含药培养基,厌氧培养24h后测pH值,并计算pH变化值(△pH值)以检测不同浓度黑蜂蜂胶对测试菌单菌生物膜状态下产酸能力的影响；⑥葸酮法测定MBEC以下的四个浓度(包括MBEC浓度)的黑蜂蜂胶对变形链球菌24h单菌生物膜产生水不溶性胞外多糖的影响。结果：①伊犁黑蜂蜂胶对变形链球菌、远缘链球菌、嗜酸乳杆菌、血链球菌、粘性放线菌、内氏放线菌的MIC分别是0.781ng·mL-1、0.39mg·mL-1、1.56mg· mL-1、0.39mg·mL-1、0.2mg·ml-1、0.2mg·mL-1; MBC分别为1.56mg·mL-11和0.78mg·mL-1、3.125mg·mL-1、0.78mg·mL-1、0.39mg·mL-1、0.39mg·mL-1;②伊犁黑蜂蜂胶对变形链球菌、远缘链球菌、血链球菌、粘性放线菌、内氏放线菌的MBEC分别是6.25mg·mL-1、1.56mg·mL-1、3.1256mg·mL-1、0.78mg·mL-1;③蜂胶可使各游离状态下的测试菌△pH降低,蜂胶各组与对照组相比差异均有统计学意义(P0.05)；④在MIC浓度时,蜂胶可使变形链球菌、粘性放线菌合成胞外多糖的能力降低,蜂胶组与对照组相比差异均有统计学意义(P0.05)；⑤伊犁黑蜂蜂胶可使各测试菌单菌生物膜△pH降低,蜂胶各组与对照组相比差异均有统计学意义(P0.05)；⑥在MBEC浓度时,蜂胶可使变形链球菌合成水不溶性胞外多糖的能力降低,蜂胶组与对照组相比差异有统计学意义(P0.05)。结论：伊犁黑蜂蜂胶对口腔主要致龋细菌及其菌斑生物膜的生长、产酸、产糖均有不同程度的抑制作用,是具有一定防龋效果的天然药物。
[Abstract]:Objective: to study the effect of Propolis on dental caries causing bacteria and its biofilm virulence factor, and to explore its anti-caries effect and possible anti-caries mechanism. Methods the minimum inhibitory concentration (MIC) of propolis against common dental caries bacteria and the minimum bactericidal concentration (MBC) were determined by liquid dilution method. The minimal growth of propolis against common dental caries was determined by the method of crystal violet staining. The membrane scavenging concentration (MBEC) 3 was configured with four concentrations below MIC (including MIC concentration) to form a medicine-containing medium with initial pH 7.0. Blank liquid medium was used as control group. The pH value was measured after anaerobic culture of bacteria solution, and pH value (pH value) was calculated to detect the effect of different concentrations of black bee propolis on acid production ability of bacteria in planktonic state. 4 the effects of four concentrations of MIC (including MIC) on Streptococcus mutans and extracellular polysaccharides produced by actinomycetes were determined by anthracanone method. The blank liquid medium was used as the control group to test the biofilm of single bacteria for 24 hours. Four concentrations of MBEC (including MBEC) were added to the medium with initial pH 7.0. After 24 hours of anaerobic culture, pH values were measured. The pH value (pH value) was calculated to detect the effect of different concentrations of Propolis on the acid-producing ability of monobacteria biofilm. 6The effects of four concentrations of MBEC (including MBEC) on the production of water-insoluble extracellular polysaccharides from the biofilm of Streptococcus mutans were determined by anthracanone method. Results the MIC of the propolis against Streptococcus mutans, Streptococcus distalis, Lactobacillus acidophilus, Streptococcus sanguis, Actinomyces glutinis and Actinomyces Neisseria were 0.781ng mL-1 0.39mg mL-1n 1.56mg mL-1n 0.39mg mL-1n 0.2mg ml-10.2mg mL-1mg respectively, and MBC were 1.56mg mL-11 and 0.78mg mL-1.3.125mg mL -1 0.78mg ml ~ (-1) 0.39mg ml ~ (-1) ~ 0.39mg mg ~ (-1) mg 路ml ~ (-1) for Streptococcus mutans, respectively. The MBEC of Streptococcus distalis, Streptococcus sanguis, Actinomyces glutinis and Actinomycetes Neoris were 6.25mg mL-1 1.56mg mL-1and 3.1256mg mL-1 0.78mg mL -1 propolis, respectively. Propolis could reduce the pH of the tested bacteria in each free state. The difference of propolis between each group and the control group was statistically significant when the MIC concentration was higher than that in the control group (P 0.05). Propolis could reduce the ability of Streptococcus mutans and actinomycetes to synthesize extracellular polysaccharides. The propolis group had significant difference from the control group. Propolis could reduce the pH of the biofilm of the tested bacteria. There was significant difference between propolis group and control group (P 0.05). Propolis could reduce the ability of Streptococcus mutans to synthesize insoluble extracellular polysaccharides when the concentration of propolis was higher than that in control group (P 0.05), and the difference between propolis group and control group was statistically significant (P 0.05). Conclusion: Yili bee propolis can inhibit the growth, acid production and sugar production of the main dental caries bacteria and their plaque biofilm in different degrees, and is a natural drug with a certain anti-caries effect.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R781.1

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