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黄连素与戊间甲酚对粪肠球菌的体外协同抗菌效果研究

龚涛 崔钰嘉 周学东 汤博钰 任彪 李雨庆

龚涛, 崔钰嘉, 周学东, 等. 黄连素与戊间甲酚对粪肠球菌的体外协同抗菌效果研究[J]. 四川大学学报(医学版), 2020, 51(6): 749-754. doi: 10.12182/20201160501
引用本文: 龚涛, 崔钰嘉, 周学东, 等. 黄连素与戊间甲酚对粪肠球菌的体外协同抗菌效果研究[J]. 四川大学学报(医学版), 2020, 51(6): 749-754. doi: 10.12182/20201160501
GONG Tao, CUI Yu-jia, ZHOU Xue-dong, et al. Synergistic Antibacterial Activity of Berberine in Combination with Amylmetacresol Against Enterococcus faecalis in vitro[J]. JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION), 2020, 51(6): 749-754. doi: 10.12182/20201160501
Citation: GONG Tao, CUI Yu-jia, ZHOU Xue-dong, et al. Synergistic Antibacterial Activity of Berberine in Combination with Amylmetacresol Against Enterococcus faecalis in vitro[J]. JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION), 2020, 51(6): 749-754. doi: 10.12182/20201160501

栏目: 口腔医学进展

黄连素与戊间甲酚对粪肠球菌的体外协同抗菌效果研究

doi: 10.12182/20201160501
基金项目: 四川大学新型冠状病毒应急项目(No.2020scunCoV-10008)资助
详细信息
    通讯作者:

    E-mail:liyuqing@scu.edu.cn

Synergistic Antibacterial Activity of Berberine in Combination with Amylmetacresol Against Enterococcus faecalis in vitro

  • 摘要:   目的  研究黄连素与戊间甲酚联合使用对粪肠球菌的抗菌作用。  方法  采用微量肉汤稀释法和活菌菌落形成单位(CFU)分别测定黄连素和戊间甲酚对粪肠球菌的最小抑菌浓度(MIC);悬液定量杀菌试验和死活菌染色检测黄连素和戊间甲酚对浮游和生物膜状态粪肠球菌的杀灭效果;扫描电子显微镜观察黄连素和戊间甲酚对粪肠球菌成熟生物膜结构的影响;CCK-8细胞增殖毒性检测试剂盒和乳酸脱氢酶细胞毒性检测试剂盒测定黄连素和戊间甲酚对人口腔黏膜上皮角质细胞(HOK)的毒性。  结果  黄连素的MIC为512 μg/mL,戊间甲酚的MIC为0.023 3%;悬液定量杀菌试验表明,512 μg/mL黄连素或0.002 33%戊间甲酚单独使用时对浮游状态粪肠球菌杀灭效果较弱,但二者联合使用时表现出协同抗菌作用;死活菌染色和扫描电子显微镜结果表明二者联合使用时对生物膜状态粪肠球菌杀灭效果较弱,但可影响生物膜结构;512 μg/mL黄连素和0.002 33%戊间甲酚单用(或联用)处理HOK细胞时,其存活率远大于损伤率,细胞毒性较低。  结论  黄连素与戊间甲酚对粪肠球菌具有协同抗菌作用,且生物安全性较好。
  • 图  1  黄连素(A)和戊间甲酚(B)的MIC测定

    Figure  1.  The MIC determination of berberine hydrochloride (BBH, A) and amylmetacresol (AMC, B)

    图  2  悬液定量杀菌试验结果

    Figure  2.  The results of quantitative suspension test

    A: The representative pictures of antibacterial effect on different drugs treatment; B: The CFU counting of antibacterial effect on different drugs treatment (n=3, *P<0.05).

    图  3  激光共聚焦显微镜观察各组死活菌比例

    Figure  3.  The ratio of living/dead bacteria observed by Laser confocal microscope

    A: Living/dead‐labeling representative images of different antibacterial drugs treating biofilm of E. faecalis (SYTO9, green showing live bacteria; PI, red showing dead bacteria; Merge showing combination of SYTO9 and PI channel); B: The fluorescence value analyzed by COMSTAT 2.1 (n=3, *P<0.05, vs. PBS).

    图  4  扫描电子显微镜观察生物膜结构。 ×8 000

    Figure  4.  The structure of biofilm observed by SEM. ×8 000

    图  5  CCK-8和LDH检测细胞毒性

    Figure  5.  The determination of cytotoxicity by CCK-8 kit and LDH assay kit

    A, D and G showed viable cells were detected after treating of different antibacterial drugs. B, E, and H showed damaged cells were detected after treating of different antibacterial drugs. C, F, and I showed cell viability and cytotoxicity were analyzed after treating of different antibacterial drugs.

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出版历程
  • 收稿日期:  2020-06-20
  • 修回日期:  2020-09-09
  • 刊出日期:  2020-11-20

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