摘要
目的观察黄连素应用后铜绿假单胞菌(PA)对环丙沙星的最低抑菌浓度(MIC)的变化和PA菌耐药基因GyrA突变情况。方法收集2021年1月至10月临床分离耐环丙沙星铜绿假单胞菌25株,测定单用或联用环丙沙星和黄连素的最低抑菌浓度(MIC),计算抑菌浓度指数(FIC),筛选出联用呈协同作用菌株,并和原临床分离耐药菌配对比较。PCR扩增PA主要耐喹诺酮的GyrA基因,并对PCR扩增产物进行测序分析。结果单用环丙沙星时,25株PA对环丙沙星的MIC均≥8 mg/L,呈耐药。单用黄连素时,MIC值为0.25~4 g/L。环丙沙星联合黄连素浓度0.125~1 g/L后,PA对环丙沙星的MIC值均有不同程度下降,下降率为100%,联用呈协同作用菌株为8株,占比32%,FIC≤0.5。联合黄连素后,黄连素应用后呈协同作用的PA菌与黄连素应用前PA菌均获得367 bp的目的片段,均有GyrA基因的表达,未发现GyrA基因突变。结论黄连素能增加环丙沙星对PA的抗菌作用,降低临床耐药PA菌的耐药性。其耐药性降低,并未引起耐药基因GyrA基因的突变逆转。
Objective To determine the changes in minimum inhibitory concentration(MIC)of Ciprofloxacin against Pseudomonas aeruginosa(PA)and the mutation in drug-resistant gene GyrA after use of Berberine.Methods Twenty-five strains of Ciprofloxacin-resistant PA clinically isolated between January 2021 and October 2021 were included in the study.The MIC against PA with Ciprofloxacin or Berberine alone or in combination was determined,respectively,and thereby the fractional inhibitory concentration(FIC)index was calculated.We sorted out the strains with synergistic response to Ciprofloxacin and Berberine combination and compared them with the original stains of drug-resistant PA.The major gene of Quinolone-resistance in PA,GyrA,was amplified by PCR,and subjected to sequencing analysis of the PCR amplification products.Results When Ciprofloxacin was used alone,its MIC against these 25 PA strains was≥8 mg/L,indicating drug resistance.When Berberine was used alone,its MIC was 0.25-4 g/L.When used in combination with Berberine at doses of 0.125-1 g/L,the MIC of Ciprofloxacin against PA showed varying decrease in 100%of the strains.There were 8 strains(32%)showing synergistic response to the combination treatment,with a Ciprofloxacin FIC≤0.5.A 367 bp target fragment was obtained from PA strains that showed synergistic response to Ciprofloxacin-Berberine combination and those without Berberine use.Either strain was expressing the GyrA gene with no mutations noted.Conclusion Berberine may increase the antibacterial effect of Ciprofloxacin on PA and reduce the drug resistance of clinical drug-resistant PA.Such attenuation in the drug resistance does not lead to mutational reversion of the drug-resistant gene GyrA.
作者
韩菊梅
肖英伦
Han Jumei;Xiao Yinglun(Department of Internal Medicine,Affiliated Hospital of Guangdong University of Technology,Guangzhou 510080,China;Department of Laboratory Medicine,Guangdong Provincial Hospital of Traditional Chinese Medicine,Guangzhou 510120,China)
出处
《中华生物医学工程杂志》
CAS
2023年第1期51-56,共6页
Chinese Journal of Biomedical Engineering
基金
广东省中医药局科研课题(20211287)。
