河北医科大学学报 ›› 2021, Vol. 42 ›› Issue (7): 759-764.doi: 10.3969/j.issn.1007-3205.2021.07.004

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芦荟苷-绿原酸混合液对变异链球菌抑制作用的体外研究

  

  1. 1.河北医科大学口腔医学院,河北医科大学口腔医院口腔预防科,河北 石家庄 050017;2.河北医科大学口腔医学院,河北医科大学口腔医院口腔病理科,河北 石家庄050017
  • 出版日期:2021-07-25 发布日期:2021-08-02
  • 作者简介:薛静秀(1991-),女,天津宝坻人,河北医科大学口腔医院医师,医学硕士,从事口腔内科疾病诊治研究。
  • 基金资助:
    河北省科技计划项目(15272703D)

Inhibitory effect of aloin-chlorogenic acid mixture on Streptococcus mutans: an in vitro study

  1. 1.School of Stomatology, Department of Stomatological Prevention, Hospital of Stomatology, Hebei 
    Medical University, Shijiazhuang 050017,China; 2.School of Stomatology, Department of 
    Oral Pathology, Hospital of Stomatology, Hebei Medical University, 
    Shijiazhuang 050017,China
  • Online:2021-07-25 Published:2021-08-02

摘要: 目的 探讨芦荟苷与绿原酸联合应用对变异链球菌生长、产酸、黏附及生物膜形成的影响。
方法 微量液体二倍稀释法分别测定芦荟苷、绿原酸对变异链球菌的最低抑菌浓度(minimum inhibitory concentration,MIC),棋盘稀释法测定芦荟苷与绿原酸混合液对变异链球菌的MIC值,通过分级抑制浓度(fractional inhibitory concentration,FIC)指数来判断两者联合应用对变异链球菌生长作用的影响,并测定低于MIC的5个浓度梯度混合液对变异链球菌产酸作用的影响;体外构建变异链球菌24 h生物膜模型,MTT法测定以上各浓度混合液作用24 h后生物膜黏附的量,并通过激光共聚焦扫描显微镜(confocal laser scanning microscope,CLSM)观察其对生物膜结构和细菌活性的影响。
结果 芦荟苷与绿原酸单独用药时,其MIC分别为0.63 g/L和5 g/L;联用的MIC为芦荟苷0.31 g/L和绿原酸2.5 g/L,FIC=1。随着混合液浓度的升高,变异链球菌菌液的△pH及OD值降低,差异有统计学意义(P<0.05)。CLSM观察显示随混合液浓度增加变异链球菌生物膜结构变稀疏,活菌数量减少。
结论 芦荟苷与绿原酸联合应用对变异链球菌的体外抗菌活性具有相加作用,较各自的单组分用药在更低浓度即可有效抑制变异链球菌的生长、产酸及黏附。


关键词: 变异链球菌, 芦荟苷, 绿原酸

Abstract: Objective To investigate the effects of a combination of aloin and chlorogenic acid on the growth, acid production, adhesion and biofilm formation of Streptococcus mutans(SM). 
Methods The minimum inhibitory concentration(MIC)  of aloin and chlorogenic acid against S.mutans were determined by liquid double micro-dilution method. MIC of mixed solution of aloin and chlorogenic acid against S.mutans was determined by the checkerboard dilution method. Fractional inhibitory concentration(FIC) index was used to evaluate the combined inhibitory effect of aloin and chlorogenic acid against S.mutans. The effect of mixed solution of aloin and chlorogenic acid, with 5 concentration gradients concentrations lower than MIC on the acid production of S.mutans, was also observed. The 24 h biofilm model of S.mutans was constructed in vitro, and MTT method was used to determine the amounts of biofilm adhesion after the above-mentioned mixture were applied for 24 h. The effects on biofilm structure and bacterial activities were observed by confocal laser scanning microscope(CLSM). 
Results When aloin and chlorogenic acid were used alone, the MIC was 0.63 g/L and 5 g/L respectively, respectively. Aloin 0.31 g/L combined with chlorogenic acid 2.5 g/L could reach MIC, with an FIC of 1. As the concentration of the mixture increased, the △pH and OD values of S.mutans was decreased(P<0.05). CLSM showed that the structure of S.mutans biofilm became thinner and the number of live bacteria decreased with the increase of the concentrations of the mixed solution.   
Conclusion The combination of aloin and chlorogenic acid has an additive effect on the in vitro antibacterial activity of S.mutans, which can effectively inhibit the growth, acid production and adhesion of S.mutans at a lower concentration than the single component.


Key words: streptococcus mutans, aloin, chlorogenic acid