盐酸小檗碱对金黄色葡萄球菌体外抗菌活性的研究

Antimicrobial activities of berberine against Staphylococcus aureus in vitro

目的探究盐酸小檗碱单药及联合应用对金黄色葡萄球菌的体外抗菌活性。方法收集首都医科大学附属北京友谊医院2020年4月至2021年3月临床分离的金黄色葡萄球菌28株,采用琼脂稀释法测定盐酸小檗碱对金黄色葡萄球菌的最低抑菌浓度,采用棋盘法检测盐酸小檗碱与左氧氟沙星、环丙沙星、甲氧苄啶、克林霉素、利福平、万古霉素和庆大霉素的联合抑菌活性,对分离株进行MLST分型,选取优势ST型(ST5,ST188,ST239)进行时间-杀菌实验和生物膜形成实验。结果盐酸小檗碱对82.1%金黄色葡萄球菌的最低抑菌浓度(MIC)为128 mg/L,10.7%菌株的MIC为64 mg/L,7.1%菌株的MIC为256 mg/L;对MRSA和MSSA的体外抑菌活性差异无统计学意义(P>0.05)。盐酸小檗碱与克林霉素、左氧氟沙星、环丙沙星、万古霉素、庆大霉素、甲氧苄啶和利福平联合应用时,分别有82.14%、78.57%、96.43%、96.43%、67.86%、17.86%和71.43%的菌株表现协同作用;盐酸小檗碱能显著抑制细菌数量和生物膜的形成,且随着药物浓度升高,抑制能力逐渐增强。结论盐酸小檗碱在体外对金黄色葡萄球菌具有良好的抑菌活性,而且与万古霉素、克林霉素、利福平、氟喹诺酮和氨基糖苷类药物具有良好的协同抑菌作用。

Objective To investigate the antimicrobial activities of berberine alone and combination with antibacterial agents against Staphylococcus aureus in vitro.Methods Twenty-eight clinical separated Staphylococcus aureus were collected from the capital medical university affiliated Beijing Friendship hospital between April 2020 to March 2021.The Multi-locus Sequence Types(MLST)and susceptibility to berberine using agar plate dilution method were tested;berberine combination with Clindamycin,levofloxacin,ciprofloxacin,vancomycin,gentamicin,trimethoprim,rifampicin were performed combining drug susceptibility test.Additionally,advantaged ST types(ST5,ST188,ST239)were selected and time-killing test and biofilm formation assay in berberine was performed.Results The Minimum Inhibitory Concentration values of Staphylococcus aureus were 128 mg/L largely(82.1%)and a minority were 64 mg/L(10.7%)and 256 mg/L(7.1%);when berberine combination with clindamycin,levofloxacin,ciprofloxacin,vancomycin,gentamicin,trimethoprim and rifampicin,82.14%,78.57%,96.43%,96.43%,67.86%,17.86%and 71.43%strains showed synergistic effects,respectively;there were not statistical difference to the antimicrobial activities against MRSA and MSSA(P>0.05);berberine showed antibacterial and anti-biofilm activities against Staphylococcus aureus,which increased with the increase of berberine concentration.Conclusion Berberine was found positive antimicrobial activities against clinical separated Staphylococcus aureus and excellent synergistic effects with vancomycin,clindamycin,rifampicin,fluoroquinolone and aminoglycoside.