积雪草酸联合亚胺培南对产KPC-2耐药肠杆菌的体外抗菌作用  

In Vitro Antibacterial Effect of Asiatic Acid Combined with Imipenem on KPC-2-producing Drug-resistant Enterobacteriaceae

作　　者：张卫 伍国强 廖远军 许竞文 邹宜诺 周永林 Zhang Wei;Wu Guoqiang;Liao Yuanjun;Xu Jingwen;Zou Yinuo;Zhou Yonglin(Liuyang Center for Animal Disease Prevention and Control,Liuyang,Hunan 410300,China;Lingling Animal Husbandry and Fisheries Affairs Center,Yongzhou,Hunan 425199,China;National Key Laboratory of Diagnosis and Treatment of Severe Zoonotic Diseases,College of Veterinary Medicine,Jilin University,Changchun,Jilin 130062,China)

机构地区：[1]浏阳市农业农村局动物疫病预防控制中心,湖南浏阳410300 [2]零陵区畜牧水产事务中心,湖南永州425199 [3]吉林大学动物医学学院,人畜共患传染病重症诊治全国重点实验室,吉林长春130062

出　　处：《中国动物检疫》2023年第8期94-102,共9页China Animal Health Inspection

基　　金：国家自然科学基金青年基金项目(32102723,32202856);中国博士后科学基金面上项目(2023M731291)。

摘　　要：碳青霉烯类抗生素是目前临床上用于治疗肠杆菌严重感染的最后防线药物。然而随着肺炎克雷伯菌碳青霉烯酶(KPCs)等的出现和流行,碳青霉烯类药物治疗广泛耐药肠杆菌(carbapenem resistant Enterobacteriaceae,CRE)感染面临巨大挑战。KPCs家族中以KPC-2为主要流行类型,其可水解几乎所有β-内酰胺类抗生素。因此针对KPC-2开展抑制剂筛选迫在眉睫。本研究通过酶活性抑制试验、抑菌试验、细菌细胞膜通透性测定和细胞毒性检测等开展了KPC-2抑制剂的筛选及其活性验证。结果显示:中药活性成分积雪草酸可显著抑制KPC-2水解β-内酰胺类抗生素的生物学活性(P <0.05,IC50=4.38μg/mL),从而可有效增强亚胺培南等碳青霉烯类抗生素的体外抑菌作用和杀菌作用。同时,积雪草酸可增强亚胺培南对细菌细胞膜的损伤作用。此外,积雪草酸(≤32μg/mL)对不同来源细胞的细胞毒性较低,这有利于进一步开发其作为碳青霉烯类抗生素佐剂并投入临床使用。综上,本研究为CRE感染治疗提供了新的可行策略和先导化合物。Carbapenem antibiotics are the last line of defense drugs used for fighting against clinically severe Enterobacteriaceae infection.However,with the emergence and prevalence of Klebsiella pneumoniae carbapenemases(KPCs),it would be a huge challenge for carbapenem drugs to fight against carbapenem resistant Enterobacteriaceae(CRE)infection.KPC-2 is most popular in KPCs family,and could hydrolyze almost allβ-lactam antibiotics.Therefore,it is urgent to screen inhibitors against KPC-2.In this study,KPC-2 inhibitor was screened and verified for its activity by enzyme activity inhibition assay,bacterial inhibition test,bacterial cell membrane permeability measurement and cytotoxicity test.The results revealed that the active ingredient of traditional Chinese medicine,asiatic acid,could significantly inhibit the biological activity of KPC-2 hydrolyzingβ-lactam antibiotics(P<0.05,IC 50=4.38μg/mL),and thus increase in vitro bacteriostasis and bactericidal action of imipenem and other carbapenem antibiotics.Meanwhile,asiatic acid could enhance the damage of imipenem to bacterial cell membrane.In addition,asiatic acid(≤32μg/mL)was with low cytotoxicity to cells from different sources,contributing to further development of its clinical use as an adjuvant for carbapenem antibiotics.In conclusion,a new feasible strategy and leading compound was provided for the treatment of CRE infection.

关 键 词：积雪草酸 KPC-2 亚胺培南 协同抗菌 

分 类 号：S855.1[农业科学—临床兽医学]