大蒜素与亚胺培南联用对耐碳青霉烯类肺炎克雷伯菌的体外抑菌效果  

In vitro antibacterial effect of allicin combined with imipenem on carbapenem-resistant Klebsiella pneumoniae

作　　者：周惠敏 马军[1] 沈进 米瑞 杨茜云 刘庆阳 王秋波[1] ZHOU Huimin;MA Jun;SHEN Jin;MI Rui;YANG Qianyun;LIU Qingyang;WANG Qiubo(Wuxi Ninth People's Hospital Affiliated to Soochow University,Wuxi,Jiangsu 214000,China)

机构地区：[1]苏州大学附属无锡九院(无锡市第九人民医院),江苏无锡214000

出　　处：《中国热带医学》2025年第1期103-109,共7页China Tropical Medicine

基　　金：江苏省卫生健康委医学科研立项面上项目(No.H2023093);江苏省卫生健康委血地寄防科研课题(No.x202319);无锡市卫生健康委中青年拔尖人才资助计划(No.BJ2023109);无锡市软科学研究课题(No.KX-24-C306)。

摘　　要：目的评价大蒜素与亚胺培南联合用药对于临床分离的耐碳青霉烯类肺炎克雷伯菌(carbapenem resistant Klebsiella pneumoniae,CRKP)的体外联合抗菌效应,为进一步动物体内药效学研究提供参考。方法根据临床药敏结果纳入22株CRKP作为实验对象。拉丝试验和碳青霉烯抑制增强试验鉴定CRKP的高黏附性及β-内酰胺酶的酶型。运用零交互效价(zero interaction potency,ZIP)方法预测大蒜素和亚胺培南的药物联合作用。使用琼脂纸片扩散法分别检测大蒜素、亚胺培南及大蒜素联合亚胺培南对CRKP的体外抗菌效果。用大蒜素和亚胺培南联合检测CRKP抑菌圈直径和最低抑菌浓度(minimal inhibitory concentration,MIC)。采用结晶紫半定量法观察大蒜素联合亚胺培南对CRKP生物膜形成的影响。结果22株CRKP中高黏附性有6株(27.27%),碳青霉烯抑制增强结果显示22株CRKP菌株均为产A类β-内酰胺酶。大蒜素对CRKP有较强的杀菌作用。根据单因素方差分析结果显示,大蒜素联合亚胺培南的抑菌圈直径(15.91±2.76)mm明显高于单独使用亚胺培南组(8.23±3.46)mm(F=46.39,P<0.001)。通过ZIP和MIC法证明大蒜素和亚胺培南联用,以相加作用为主。利用单因素方差分析结晶紫吸光度值,与对照组(0.213±0.056)相比,大蒜素联合亚胺培南组CRKP生物膜总量(0.134±0.045)降低(F=3.211,P=0.045)。结论体外抑菌试验结果显示,大蒜素与亚胺培南联用后能显著提高亚胺培南对CRKP的抑菌能力,并可有效抑制生物膜形成。Objective To evaluate the in vitro synergistic antimicrobial effect of allicin combined with imipenem on clinically isolated carbapenem-resistant Klebsiella pneumoniae(CRKP),providing a reference for further in vivo pharmacodynamic studies.Methods Twenty-two strains of CRKP were selected as experimental subjects based on clinical drug sensitivity results.Pulling assay together with carbapenem inhibition enhancement assay were leveraged to identify the high adherence andβ-lactamase phenotypes of CRKP.The combined effect of allicin and imipenem was confirmed using the zero interaction potency(ZIP)method.The antimicrobial effects of allicin,imipenem,and their combination against CRKP were tested separately in vitro by agar paper diffusion method.The organic combination of allicin and imipenem was exploited for detecting the diameter of the CRKP inhibitory circle and minimal inhibitory concentration(MIC).The crystal violet semi-quantitative method was utilized to observe the effect of allicin combined with imipenem on CRKP biofilm formation.Results Among the 22 strains of CRKP,6 strains exhibited high adhesion,accounting for 27.27%.The results of carbapenem inhibition enhancement demonstrated that all strains of CRKP were class Aβ-lactamase-producing.Allicin had a strong bactericidal effect on CRKP.According to the results of one-way ANOVA,the diameter of the circle of inhibition of allicin combined with imipenem(15.91±2.76)mm was significantly higher than that of the imipenem alone group(8.23±3.46)mm(F=46.39,P<0.001).The ZIP scoring and MIC methods confirmed that the combination of allicin and imipenem primarily exerted an additive effect.One-way ANOVA analysis of crystal violet absorbance values showed that compared to the control group(0.213±0.056),the total CRKP biofilm amount in the allicin combined with imipenem group was reduced(0.134±0.045)(F=3.211,P=0.045).Conclusions The results of bacterial inhibition experiments in vitro show that the combination of allicin and imipenem significantly increases the i

关 键 词：大蒜素 亚胺培南 肺炎克雷伯菌 联合药敏 生物膜 

分 类 号：R515[医药卫生—内科学]