摘要
目的:研究痰热清与临床常用抗生素对多重耐药铜绿假单胞菌(multidrug resistant Pseudomonas aeruginosa,MDR-PA)的相互作用,以及对细菌外排泵的作用研究。方法:利用抗生素对细菌进行药敏实验。纸片扩散法(Kirby-Bauer,K-B)结合外排泵抑制剂(50 mg·L^-1)检测抑菌圈直径以及实时荧光定量聚合酶链式反应(Real-time PCR)检测外排泵基因表达量筛选外排泵阳性菌株。KB法观察痰热清(终质量浓度3 g·L^-1)与抗生素对抑菌圈直径的变化。菌株与痰热清、抗生素亚抑菌浓度共培养进行Real-time PCR检测。KB法观察痰热清持续作用外排泵阳性菌后对抑菌环直径的影响。结果:经鉴定筛选得到两株MDR-PA外排泵阳性菌株。痰热清与阿洛西林、氨曲南、美罗培南、头孢他啶、头孢哌酮、舒普深均有协同抗菌作用。在抑制细菌外排泵基因表达水平方面,4种药物按作用效果强弱比较为头孢哌酮>痰热清>头孢他啶>舒普深。痰热清持续作用外排泵阳性菌株后与头孢他啶、头孢哌酮、舒普深联合用药疗效较好。结论:痰热清与头孢他啶,头孢哌酮,舒普深可通过下调细菌外排泵基因表达,使细菌耐药性减弱,降低抗生素临床使用剂量,从而起到抑菌作用。
Objective:To study the interaction between Tanreqing injection and commonly used antibiotics against multi-drug resistant Pseudomonas aeruginosa(MDR-PA)and the effect on bacterial efflux pump.Method:Antibiotic susceptibility test was performed with bacteria.Paper diffusion method(KirbyBauer,KB)combined with efflux pump inhibitor(50 mg·L^-1)was used to measure the diameter of the inhibition zone,and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect gene expression of efflux pump Positive efflux pump strain.KB method was used to observe the changes of Tanreqing(final concentration 3 g·L^-1)and antibiotics on the diameter of the zone of inhibition.Strains were co-cultured with Tanreqing and antibiotic sub-inhibitory concentrations for Real-time PCR detection.KB method was used to observe the effect of Tanreqing on the diameter of bacteriostatic ring after the continuous use of efflux pump-positive bacteria.Result:Two MDR-PA efflux pump-positive strains were identified and screened.Tanreqing has synergistic antibacterial effect with aloxicillin,aztreonam,meropenem,ceftazidime,cefoperazone,and Shupushen.In inhibiting the expression levels of bacterial efflux pump genes,the four drugs were compared by the effect:cefoperazone>Tanreqing>ceftazidime>Shupushen.After Tanreqing continued to act on efflux pump-positive strains,it could have a better effect in combination with ceftazidime,cefoperazone,and Shupushen.Conclusion:Tanreqing,ceftazidime,cefoperazone,and Shupushen can reduce the drug resistance of bacteria by down-regulating the expressions of bacterial efflux pump genes,and reducing the clinical dose of antibiotics,and thus play a bacteriostatic effect.
作者
李悦怡
李贤煜
杨伟峰
王毅
何忠梅
LI Yue-yi;LI Xian-yu;YANG Wei-feng;WANG Yi;HE Zhong-mei(Jilin Agricultural University,Changchun 130000,China;China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第14期92-98,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
财政部基本科研业务项目(Y201701)。
作者简介
第一作者:李悦怡,在读硕士,从事中药抗菌及中药药理研究工作,E-mail:1668710281@qq.com;通信作者:王毅,研究员,博士生导师,从事中西医微生物免疫学研究工作,Tel:010-64019508,E-mail:wangyi02@tsinghua.org.cn;通信作者:何忠梅,教授,硕士研究生导师,从事中药有效成分、作用机制研究及产品开发工作,E-mail:heather78@126.com。
