黄连素与戊间甲酚对粪肠球菌的体外协同抗菌效果研究  

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

作　　者：龚涛[1] 崔钰嘉 周学东[1] 汤博钰 任彪[1] 李雨庆[1] GONG Tao;CUI Yu-jia;ZHOU Xue-dong;TANG Bo-yu;REN Biao;LI Yu-qing(State Key Laboratory of Oral Diseases,National Clinical Research Center for Oral Diseases,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四川大学华西口腔医院,成都610041

出　　处：《四川大学学报（医学版）》2020年第6期749-754,共6页Journal of Sichuan University(Medical Sciences)

基　　金：四川大学新型冠状病毒应急项目(No.2020scunCoV-10008)资助。

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512μg/mL,戊间甲酚的MIC为0.0233%;悬液定量杀菌试验表明,512μg/mL黄连素或0.00233%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;512μg/mL黄连素和0.00233%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.Objective To study the antibacterial effect of berberine combined with amylmetacresol on Enterococcus faecalis. Methods Both dilution method and live bacteria CFU were used to determine the minimum inhibitory concentration(MIC) of berberine and amylmetacresol on E. faecalis. The killing effect of berberine and amylmetacresol on planktonic E. faecalis was detected by suspension quantitative germicidal test and live/dead bacteria staining. The effects of berberine and amylmetacresol on the structure of mature biofilm of E. faecalis was observed by scanning electron microscopy(SEM). The toxicity of berberine and amylmetacresol on human oral keratinocytes(HOK)was determined by CCK-8 cell proliferation and cytotoxicity assay and cytotoxicity LDH assay. Results The MIC of berberine was 512 μg/mL, and the MIC of amylmetacresol was 0.023 3%. 512 μg/mL berberine and 0.002 33%amylmetacresol had a weak killing effect on planktonic E. faecalis alone, while they showed a synergistic antibacterial effect in combination. Cell survival in the biofilm was only slightly changed by berberine and amylmetacresol. The structure of biofilm was obviously changed by berberine and amylmetacresol. 512 μg/mL berberine and 0.002 33%amylmetacresol alone or in combination showed the survival rate was much higher than the injury rate, suggesting berberine and amylmetacresol had a low cytotoxicity. Conclusion Berberine and amylmetacresol had synergism against E. faecalis, and the biological safety of the combination use was better.

关 键 词：黄连素 戊间甲酚 粪肠球菌 协同抗菌 

分 类 号：R96[医药卫生—药理学]