积雪草酸抑制变异链球菌生物膜形成及致龋能力的研究  

Anti-cariogenic and Anti-biofilm Effect of Asiatic Acid on Streptococcus Mutans

作　　者：税钰森 杨燃[2] SHUI Yusen;YANG Ran(State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University. Chengdu 610041, China;State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Pediatric Dentistry, West China Hospital of Stomatology, Sichuan University. Chengdu 610041, China)

机构地区：[1]口腔疾病研究国家重点实验室,国家口腔疾病临床医学研究中心,华西口腔医学院(四川大学),四川成都610041 [2]口腔疾病研究国家重点实验室,国家口腔疾病临床医学研究中心,华西口腔医院儿童口腔科(四川大学),四川成都610041

出　　处：《口腔医学研究》2021年第5期401-406,共6页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:81800989)。

摘　　要：目的:研究积雪草酸对变异链球菌(Streptococcus mutans,S.mutans)生物膜的抑制作用及对其产酸和产胞外多糖(extracellular polysaccharides,EPS)的影响。方法:通过药敏试验测定积雪草酸对S.mutans浮游菌的最低抑菌浓度(Minimum inhibitory concentration,MIC);结晶紫试验和MTT试验分别探究积雪草酸对S.mutans生物膜的总量和代谢活性的影响;采用蒽酮-硫酸法,场发射扫描电镜(Field emission scanning electron microscopy,FESEM)和激光共聚焦显微镜(Confocal laser scanning microscopy,CLSM)测定积雪草酸对S.mutans生物膜EPS的产生和结构形貌的影响;pH计测定积雪草酸对S.mutans产酸的影响;CCK-8法检测积雪草酸的细胞毒性。结果:积雪草酸对S.mutans的最低抑菌浓度为20μg/mL,在亚抑菌浓度(10μg/mL和5μg/mL)下均能有效抑制S.mutans生物膜形成以及产酸和产EPS。同时,积雪草酸无明显的细胞毒性。结论:积雪草酸在一定浓度下能有效抑制S.mutans生物膜形成,并能降低其产酸和产EPS等致龋毒力。Objective:To analyze the inhibitory effect of asiatic acid on Streptococcus mutans(S.mutans)biofim formation and its acid and extracellular polysaccharides(EPS)production.Methods:The planktonic bacterial susceptibility assay was used to determine the minimum inhibitory concentration(MIC)of asiatic acid on S.mutans.Crystal violet assay and MTT assay were performed to determine the effect of asiatic acid on the biomass and metabolic activity of S.mutans biofilm.Anthrone-sulfuric acid method,field emission scanning electron microscopy(FESEM),and confocal laser scanning microscopy(CLSM)were performed to observe the biofilm morphology and EPS synthesis after treatment by asiatic acid.The effect of asiatic acid on the acid production of S.mutans was performed by pH drop assay.CCK-8 kit assay was conducted to assess the cytotoxicity of asiatic acid.Results:The MIC of asiatic acid on S.mutans was 20μg/mL.The sub-MICs(10μg/mL and 5μg/mL)were all effective on the reduction of the biofilm formation,acid production and EPS synthesis of S.mutans.No significant cytotoxicity of asiatic acid was found.Conclusion:Asiatic acid is effective on the inhibition of S.mutans biofilm formation,its acid production,and EPS synthesis.

关 键 词：积雪草酸 变异链球菌 生物膜 

分 类 号：R285[医药卫生—中药学]