Effect of Tanreqing Injection Combined with Imipenem-cilastatin in Inhibiting Extensively-drug Resistant Pseudomonas aeruginosa

Ya-jun XING; Wei-feng YANG; Ya-nan SUN; Shu-hua MA; Tong-tong JIANG; Gai-ying HE; Xiao-li ZHANG; Yi WANG

doi:10.13422/j.cnki.syfjx.20191107

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Effect of Tanreqing Injection Combined with Imipenem-cilastatin in Inhibiting Extensively-drug Resistant Pseudomonas aeruginosa

Pharmacology | 更新时间：2021-02-09

- Effect of Tanreqing Injection Combined with Imipenem-cilastatin in Inhibiting Extensively-drug Resistant Pseudomonas aeruginosa
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 11, Pages: 60-66(2019)
- 作者机构：
  
  1.中国中医科学院　医学实验中心，北京　 100700；
  2.吉林农业大学，长春　 130118；
  3.上海凯宝药业股份有限公司，上海　 201401
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191107
  CLC： R2-0; R22; R285.5
- Received：07 November 2018，
  
  Published：05 June 2019
- 稿件说明：
移动端阅览
Ya-jun XING, Wei-feng YANG, Ya-nan SUN, et al. Effect of Tanreqing Injection Combined with Imipenem-cilastatin in Inhibiting Extensively-drug Resistant Pseudomonas aeruginosa[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(11): 60-66.
DOI：

Ya-jun XING, Wei-feng YANG, Ya-nan SUN, et al. Effect of Tanreqing Injection Combined with Imipenem-cilastatin in Inhibiting Extensively-drug Resistant Pseudomonas aeruginosa[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(11): 60-66. DOI： 10.13422/j.cnki.syfjx.20191107.

摘要

目的：

观察痰热清注射液(痰热清)与亚胺培南西司他丁联合对广泛耐药铜绿假单胞菌的抑菌作用，并研究其作用机制，为临床治疗耐药菌提供参考。

方法：

将医院临床分离到广泛耐药铜绿假单胞菌采用微量稀释法检测痰热清与亚胺培南西司他丁最小抑菌浓度(MIC)；棋盘法检测联合用药抑菌效果；微孔板法检测对成熟生物膜内细菌代谢活性的影响；BioFlux动态观察药物对生物膜的影响并用激光共聚焦扫描显微镜观察活死细菌的分布；扫描电子显微镜(SEM)观察药物作用后细菌的形态变化。

结果：

亚胺培南西司他丁和痰热清的MIC分别为512 mg·L

－1

和

16 500 mg·L

－1

。棋盘法结果显示痰热清可增强亚胺培南西司他丁的敏感性，二者协同抑菌，并确定联合用药浓度。与空白组或单用亚胺培南西司他丁组比较，联合用药各组显著减少生物膜内活菌量(

0.05，

0.01)。BioFlux结果表明与空白组或单用药物组比较，联合用药各组可破坏生物膜结构，减小生物膜面积(

0.05)；荧光染色结果显示联合用药对动态生物膜内细菌的代谢活性有明显抑制。SEM显示痰热清可抑制细菌分裂。

结论：

痰热清与亚胺培南西司他丁联合对浮游和生物膜状态的细菌生长有抑制作用，还可对生物膜产生破坏作用。

Abstract

Objective:

To observe the effect of combination of Tanreqing injection(Tanreqing) and imipenem-cilastatin on extensively-drug resistant

Pseudomonas aeruginosa

(XDPA)

and study the mechanism of the combination.

Method:

The minimum inhibitory concentrations (MICs) of Tanreqing and imipenem-cilastatin against planktonic XDPA strain isolated in clinic were determined by the broth microdilution method. The checkerboard method was used to evaluate the combination effect. The bacterial metabolic activity in mature biofilm was studied by microtiter-plate test. The destructive effect of combination drugs on dynamic biofilm was observed by using BioFlux system

and viable cells were examined by confocal laser scanning microscope (CLSM) after treatment. The scanning electron microscopy (SEM) was used for observing

Pseudomonas aeruginosa

and length measurement.

Result:

The MIC values of imipenem-cilastatin and Tanreqing were 512 mg·L

-1

and more than 16 500 mg·L

-1

. The checkerboard analysis showed that Tanreqing could enhance the sensitivity of imipenem-cilastatin

while the combination drugs synergistically inhibited the growth of bacteria. Compared with the control group or the imipenem-cilastatin individual group

the combined drugs significantly reduced the amount of living bacteria in the biofilm (

0.05

0.01). BioFlux results showed that the combination drugs destructed the biofilm structure and reduced the area coverage (

0.05) by comparing with the control group or the single drug group. The results of fluorescent staining showed that the combination drug significantly inhibited the metabolic activity of bacteria in dynamic biofilm. Tanreqing inhibited bacterial division to achieve the antibacterial effect.

Conclusion:

Tanreqing and imipenem-cilastatin synergistically inhibit the bacterial growth in planktonic and biofilm states

and destruct biofilms.

关键词

Keywords

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