抗变异链球菌卵黄抗体对变异链球菌致龋毒力因子及菌斑生物膜的影响  

Effect of anti-Streptococcus Mutans Yolk Antibody on Virulence Factors and Plaque Biofilm induced by Streptococcus Mutans

作　　者：周莉莉 徐燕[1] 蒋鹏 徐涵颖 辛保见 沈继龙[2] 程婷[3] ZHOU Li-li;XU Yan;JIANG Peng;XU HAN-ying;XIN Bao-jian;SHEN Ji-long;CHENG Ting(College & Hospital of Stomatology,Anhui Medical University,Key Lab of Oral Diseases Research of Anhui Province,Hefei 230032,China;Key Lab of Zoonoses of Anhui Province,Anhui Medical University,Hefei 230032,China;Anhui Anke Biotechnology (Group) Co.Ltd,Hefei 230032,China.)

机构地区：[1]安徽医科大学口腔医学院安徽医科大学附属口腔医院安徽省口腔疾病研究重点实验室,安徽合肥230032 [2]安徽医科大学人兽共患病安徽省重点实验室,安徽合肥230032 [3]安徽安科生物工程(集团)股份有限公司,安徽合肥230032

出　　处：《口腔医学研究》2019年第5期448-452,共5页Journal of Oral Science Research

基　　金：安徽省教育厅自然科学重大项目(编号:KJ2017ZD17)

摘　　要：目的:制备并纯化抗变异链球菌卵黄抗体(抗S.mutans-IgY),观察其对变异链球菌(Streptococcus mutans,S.mutans)胞外多糖、葡糖基转移酶活性、生物膜形成及活性的影响。方法:将甲醛灭活的S.mutans免疫150日龄白航蛋鸡4次,每次间隔10 d,取鸡蛋用两步硫酸铵沉淀法提取IgY,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测免疫后效价变化。蒽酮法测定水不溶性及水溶性胞外多糖;Neson-Somogyi法测定葡萄糖基转移酶活性;二甲氧唑黄[2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrzo lium hydroxide,XTT]还原比色法研究IgY对生物膜总量的影响;采用激光共聚焦显微镜观察牙菌斑生物膜中死菌和活菌的构成。结果:经4次免疫后抗体最高效价为1∶256000,维持40 d;IgY对S.mutans合成水不溶性胞外多糖的能力较对照组有明显抑制(P<0.01);药物作用后S.mutans还原糖、酶活力均降低(P<0.01);XTT法结果显示IgY对S.mutans生物膜抑制率为17.02%～74.13%;激光共聚焦显微镜观察到随着药液浓度增加,绿色的活菌、团块结构减少,生物膜厚度明显减小。结论:一定浓度IgY能够抑制S.mutans的增殖,具有抗龋作用。Objective:The anti-Streptococcus mutans egg yolk immunoglobulin(anti-S.mutans-IgY)was prepared and purified,and its effect on the extracellular polysaccharides capacity,glucosyltransferase activity,biofilm formation and activity of S.mutans was observed.Methods:Ten 150 day-old laying hens were immunized with S.mutans and IgY was isolated from the egg yolk by the water dilution(WD)method and 2 steps of ammonium sulfate purification.ELISA was used to detect changes in titer after immunization.The anthrone method was used to observe the water soluble glucan(WSG)and water insoluble glucan(WIG),the Neson-Somogyi method was used to determine the activity of GTF enzyme,and the XTT method was used to study the effect of IgY on the total amount of biofilm.Results:After 4 immunizations,the highest titer of the antibody was 1:256000,which was maintained for 40 days.IgY inhibited the WIG synthesis of S.mutans,and decreased the S.mutans reducing sugar and enzyme activity(P<0.01).The XTT method showed that the inhibition rate of IgY to S.mutans biofilm was 17.02%-74.13%.Microscopy showed that the green viable bacteria and structure decreased and the biofilm formation was significantly inhibited as the concentration of the drug solution increased.Conclusion:IgY can inhibit the proliferation of S.mutans and has an anticaries effect.

关 键 词：变异链球菌 卵黄抗体 葡糖基转移酶 水不溶性多糖 菌斑生物膜 

分 类 号：R781.1[医药卫生—口腔医学]