槲皮素对变形链球菌生物膜形成的影响  

Inhibitory effect of quercetin on the biofilm formation of Streptococcus mutans

作　　者：岳嘉曦 杨宏业[1] 韩林 朱敏英[3] 宋芳芳[1] 黄翠[4] 

机构地区：[1]武汉大学口腔医学院口腔基础医学省部共建国家重点实验室培育基地和口腔生物医学教育部重点实验室,430079 [2]华润武钢总医院皮肤科,武汉430080 [3]湖北民族学院中医药学院中药专业教研室,湖北恩施445000 [4]武汉大学口腔医学院修复科,430079

出　　处：《中华口腔医学杂志》2016年第6期368-373,共6页Chinese Journal of Stomatology

基　　金：国家自然科学基金（81371191）

摘　　要：目的 观察植物提取物槲皮素对变形链球菌生物膜的抑制作用,初步探寻其作用机制和生物相容性,为槲皮素在口腔生物材料中的应用提供理论依据.方法 将槲皮素储存液稀释后加入变形链球菌培养基,结晶紫染色法测定生物膜总量;后续检测根据槲皮素质量浓度分为3组：对照组（0 mg/L）、200 mg/L槲皮素组和400 mg/L槲皮素组,激光扫描共聚焦显微镜观察槲皮素处理下的变形链球菌生物膜形态和活菌、死菌比例;实时定量荧光PCR检测变形链球菌相关基因表达量;甲基噻唑基四唑法检测槲皮素对人牙髓细胞的毒性作用.结果 槲皮素可以显著抑制变形链球菌生物膜的形成,抑制率达（86.16±0.45）％,并能有效清除成熟生物膜,清除率为（43.04±0.53）％,与对照组相比差异均有统计学意义（P=0.00）;实验组24 h后变形链球菌致密生物膜结构被破坏,与对照组相比gtfB、gtfC、comD和comE基因表达量降低均超过50％,差异均有统计学意义（P＜0.05）,而luxS基因表达量在4h和24 h分别升高至（2.18±0.24）和（2.84±0.26）,均显著高于对照组（P＜0.05）;槲皮素对人牙髓细胞的毒性作用不明显.结论 槲皮素可以通过抑制相关基因表达有效减少变形链球菌生物膜的形成,并具有良好的牙髓细胞相容性,在口腔生物材料领域有广阔的应用前景.Objective To observe the inhibitory effect of quercetin on the biofilm formation of Streptococcus mutans（Sm）,to preliminarily reveal the possible underlying mechanisms,and to evaluate the cytotoxicity of quercetion to human dental pulp cells so as to provide the theoretical basis for the application of quercetin in oral biomaterials.Methods Quercetin storage solution was diluted to 0,3.125,6.25,12.5,25,50,100,200,400 and 800 mg/L,and added into Sm medium for 4 h and 24 h,crystal violet staining was used to evaluate the biofilm volume.In subsequent detections,three groups were set：control（0 mg/L）,200 mg/L quercetin and 400 mg/L quercetin.Confocal laser scanning microscopy was used to observe the morphology of the biofilm;qPCR for gtfB,gtfC,comD,comE,and luxS were assessed to preliminarily investigate the mechanisms.Finally,the methyl thiazolyl tetrazolium（MTT） test using human dental pulp cells was used to investigate cytotoxicity.Results Quercetin could significantly inhibit up to （86.16±0.45）％ of the biofilm formation of Sm （Compared with the control group P=0.00） and effectively removed （43.04±0.53）％ of the mature biofilm（Compared with the control group P=0.00）.Confocal laser scanning microscopy photographs showed that after co-incubated for 24 h,the dense biofilm structures of the experimental group were destroyed by quercetin both at 200 mg/L and 400 mg/L.Quercetin suppressedover 50％ of the expression of gtfB,gtfC,comD,comE（compared with the control group P＜0.05） and promoted the expression of luxS up to 2.18 ±0.24 and 2.84±0.26 after 4 h and 24 h,respectively（compared with the control group P＜0.05）.Quercetin also exhibited acceptable compatibility for human dental pulp cells.Conclusions Quercetin could effectively reduce the biofilm formation of Sm by inhibiting the expression of the related genes,and exhibited no cytotoxicity for human dental pulp cells.Quercetin has good potential to be applied in oral biological materials.

关 键 词：槲皮素 链球菌 变异 生物膜 显微镜检查 共焦 

分 类 号：R781.1[医药卫生—口腔医学]