人参皂苷Rh2型联合氟化钠对变异链球菌生长及生物膜形成的抑制作用  

Inhibitory effects of ginsenoside Rh2 combined with sodium fluoride on growth and biofilm formation of Streptococcus mutans

作　　者：方圆 刘晓璇 王瑞[1,2] FANG Yuan;LIU Xiaoxuan;WANG Rui(Department of Stomatology Prevention,Stomatology Hospital,Jilin University,Changchun 130021,China;Key Laboratory of Tooth Development and Jaw Bone Remodeling and Regeneration,Jilin Province,Changchun 130021,China)

机构地区：[1]吉林大学口腔医院口腔预防科,吉林长春130021 [2]吉林省牙发育及颌骨重塑与再生重点实验室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2022年第2期407-413,共7页Journal of Jilin University：Medicine Edition

基　　金：吉林省卫计委卫生计生科技项目(2017W001);吉林省卫计委卫生适宜技术扶贫项目(2017FP024);吉林省卫健委卫生健康研究项目(2020JK001)。

摘　　要：目的:探讨人参皂苷Rh2型(G-Rh2)联合氟化钠(NaF)对体外变异链球菌生长及生物膜形成的抑制作用,为其将来在口腔临床上的应用提供理论依据。方法:采用微量稀释法分别测量G-Rh2与NaF对变异链球菌的半数最小抑制浓度(MIC_(50));采用棋盘微量稀释法测量G-Rh2与NaF联合应用于变异链球菌的MIC_(50)值,并计算分级抑菌浓度(FIC)指数来判断两者的联合效应,当FIC<0.5时证明2种药物间具有协同抑制变异链球菌的作用;采用结晶紫染色实验分别测量G-Rh2与NaF对变异链球菌的半数最小生物膜抑制浓度(MBIC50)值;实验分为空白对照组、MIC_(50)G-Rh2组、MIC_(50)NaF组和MIC_(50)G-Rh2+MIC_(50)NaF联合组,采用生长曲线及产酸实验评估各组变异链球菌浮游菌生长速度及产酸抑制率;实验分为空白对照组、MBIC50G-Rh2组、MBIC50NaF组及MBIC50G-Rh2+MBIC50NaF联合组,采用结晶紫染色实验及扫描电镜观察各组变异链球菌生物膜形成量及结构的变化;空白对照组内不含任何药物。结果:G-Rh2与NaF作用于变异链球菌的MIC_(50)分别为25.000和125.000 mg·L^(-1);G-Rh2与NaF联合应用时MIC_(50)分别为5.00和31.25 mg·L^(-1),FIC指数为0.45,小于0.50,证明2种药物间具有协同抑制变异链球菌的作用;G-Rh2与NaF作用于变异链球菌的MBIC50分别为22.5和62.5 mg·L^(-1)。与空白对照组、MIC_(50)G-Rh2组和MIC_(50)NaF组比较,MIC_(50)G-Rh2+MIC_(50)NaF联合组变异链球菌的生长速度明显降低(P<0.05)。与空白对照组比较,MIC_(50)G-Rh2组、MIC_(50)NaF组和MIC_(50)G-Rh2+MIC_(50)NaF联合组ΔpH值均降低,且MIC_(50)G-Rh2+MIC_(50)NaF联合组ΔpH值低于MIC_(50)G-Rh2组和MIC_(50)NaF组(P<0.05);MIC_(50)G-Rh2+MIC_(50)NaF联合组的产酸抑制率高于MIC_(50)G-Rh2组和MIC_(50)NaF组(P<0.05)。MBIC50G-Rh2+MBIC50NaF联合组的生物膜形成量明显少于空白对照组、MBIC50G-Rh2组和MBIC50NaF组(P<0.01)。扫描电镜观察,MBIC50G-Rh2组和MBIC50NaF组生物膜厚度降低,�Objective:To explore the inhibitory effects of ginsenoside Rh2(G-Rh2)combined with sodium fluoride(NaF)on the growth of Streptococcus mutans and biofilm formation in vitro,and to provide theoretical basis for their application in stomatological clinic.Methods:The half minimum inhibitory concentrations(MIC_(50))of G-Rh2 and NaF against Streptococcus mutans were measured by microdilution method;the MIC_(50)value of G-Rh2 combined with NaF on Streptococcus mutans was measured by chessboard microdilution method,and the fractional inhibitory concentration(FIC)value was calculated to judge the combined effect of these two drugs.When FIC<0.5,it was proved that the two drugs had synergistic inhibitory effect on Streptococcus mutans.The half minimum biofilm inhibition concentrations(MBIC;)of G-Rh2 and NaF against Streptococcus mutans were measured by crystal violet staining test.The experiment was divided into blank control group,MIC_(50)G-Rh2 group,MIC_(50)NaF group and MIC_(50)G-Rh2+MIC_(50)NaF combined group,the growth curve and acid production test were used to evaluate the growth speeds and inhibitory rates of acid production of Streptococcus mutans in each group;the experiment was divided into blank control group,MBIC50G-Rh2 group,MBIC50NaF group and MBIC50G-Rh2+MBIC50NaF combined group,the changes of biofilm formation of Streptococcus mutans and structures were observed by crystal violet staining test and scanning electron microscope in each group.The blank control group did not contain any drugs.Results:The MIC_(50)of G-Rh2 and NaF acting on Streptococcus mutans alone were 25.000 and 125.000 mg·L^(-1),respectively;the MIC_(50)of G-Rh2 combined with NaF on Streptococcus mutans were 5.00 and 31.25 mg·L^(-1),the FIC index was 0.45 and less than0.5,which proved that the two drugs had synergistic inhibitory effect on Streptococcus mutans.The MBIC50of G-Rh2 and NaF were 22.5 and 62.5 mg·L^(-1).The growth speed of Streptococcus mutans in MIC_(50)G-Rh2+MIC_(50)NaF combined group was significantly lower than those in bl

关 键 词：变异链球菌 人参皂苷Rh2型 氟化钠 联合药敏实验 

分 类 号：R781.1[医药卫生—口腔医学]