松油烯-4-醇与α-红没药醇对金黄色葡萄球菌和痤疮丙酸杆菌协同抑菌作用研究

SYNERGISTIC BACTERIOSTATIC EFFECT OF TERPINEN-4-OL ANDα-BISABOLOL ON STAPHYLOCOCCUS AUREUS AND CUTIBACTERIUM ACNES

目的探讨松油烯-4-醇(T4O)和α-红没药醇(Bis)对于金黄色葡萄球菌(S.aureus)和痤疮丙酸杆菌(C.acnes)的协同抑菌作用。方法采用微量棋盘稀释法测定T4O、Bis联用时上述两种实验菌的最小抑菌浓度(MIC),并计算部分抑菌浓度指数(FICI)。对两种实验菌各设置MIC组(加入MIC的T4O、Bis)、2×MIC组(加入2倍MIC的T4O、Bis)及阴性对照组(不加T4O、Bis),绘制T4O、Bis联用时两种实验菌各组的时间-杀菌曲线,测定联用时两种实验菌各组的菌株表面Zeta电位(ZP)、细胞内核酸及蛋白质泄漏情况、细菌生物膜破坏情况以及S.aureus的呼吸链脱氢酶活性,透射电镜观察联用对两种实验菌各组菌株形态的影响。结果T4O和Bis联用时S.aureus及C.acnes的FICI<0.5,其中T4O对S.aureus的MIC为0.62 g/L,对C.acnes的MIC为0.31 g/L;时间-杀菌曲线显示,联用可明显抑制两种细菌生长。与阴性对照组相比,联用时两种细菌的ZP均降低,细菌细胞内核酸、蛋白质泄漏增加,菌体丧失正常形态,细菌生物膜被破坏,且S.aureus呼吸链脱氢酶活力降低。结论T4O和Bis对S.aureus及C.acnes具有协同抑菌作用,两者联合可能成为抑菌产品的有效成分。

Objective To investigate the synergistic bacteriostatic effect of terpinen-4-ol(T4O)andα-bisabolol(Bis)on Staphylococcus aureus(S.aureus)and Cutibacterium acnes(C.acnes).Methods The microdilution checkerboard technique was used to determine the minimum inhibitory concentration(MIC)of T4O combined with Bis on the two experimental bacteria,and fractional inhibitory concentration index(FICI)was calculated.An MIC group(T4O and Bis at a concentration of MIC),a 2×MIC group(T4O and Bis at a concentration of 2×MIC),and a negative control group(no T4O or Bis added)were set up for each experimental bacterium;the time-bactericidal curve was plotted for each experimental bacterium after the addition of both T4O and Bis;each group of the two experimental bacteria was measured in terms of Zeta potential(ZP)on the surface of the strain,leakage of intracellular nucleic acid and protein,damage of bacterial biofilm,and respiratory chain dehydrogenase activity in S.aureus after the addition of both T4O and Bis;a transmission electron microscope was used to observe the influence of T4O combined with Bis on the morphology of the two experimental bacteria.Results The combination of T4O and Bis for S.aureus and C.acnes had an FICI of<0.5,and T4O had an MIC of 0.31 g/L for C.acnes and 0.62 g/L for S.aureus.The time-bactericidal curve showed that the combination of T4O and Bis significantly inhibited the growth of both bacteria.Compared with the negative control group,the combination group had a significant reduction in ZP,a significant increase in the leakage of intracellular nucleic acid and protein,loss of the normal morphology of bacterial body,damage of bacterial biofilm,and a reduction in respiratory chain dehydrogenase activity in S.aureus.Conclusion T4O and Bis have a synergistic bacteriostatic effect on S.aureus and C.acnes,and the combination of T4O and Bis may become an effective component of antibacterial products.