黄芩对安普霉素耐药大肠杆菌的耐药消除研究

doi:10.16431/j.cnki.1671-7236.2023.03.042

摘要/Abstract

摘要： 【目的】探讨黄芩对安普霉素耐药大肠杆菌的耐药消除作用。【方法】制备黄芩醇提物,采用琼脂稀释法检测黄芩醇提物对耐药大肠杆菌的最小抑菌浓度(MIC),通过耐药消除试验和影印法检测耐药消除率。进而采用微量肉汤稀释法和纸片扩散法检测消除前后大肠杆菌耐药性的变化,扫描电镜法观察细菌形态的变化,实时荧光定量PCR法检测耐药相关基因相对表达量的变化。【结果】黄芩醇提物对安普霉素耐药大肠杆菌的MIC为41.25 mg/mL,对耐药大肠杆菌的耐药消除率为11.7%~17.0%。黄芩醇提物处理后,大肠杆菌的MIC值由256 μg/mL降至16~32 μg/mL;扫描电镜观察发现,黄芩醇提物处理组细菌表面受损,菌体形态异常;实时荧光定量PCR检测结果显示,黄芩醇提物极显著提高了大肠杆菌中ompF、fimA、phnE和fhuF基因mRNA的相对表达量(P＜0.01)。【结论】黄芩醇提物能提高ompF、fimA、phnE和fhuF基因的表达水平,破坏细菌表面,提高大肠杆菌敏感性,发挥对安普霉素耐药大肠杆菌的耐药消除作用。

关键词: 黄芩; 大肠杆菌; 耐药消除; 安普霉素

Abstract: 【Objective】 This study was aimed to explore the drug-resistance elimination of Scutellaria baicalensis (S.baicalensis) on apramycin-resistant Escherichia coli (E.coli).【Method】 The alcohol extract of S.baicalensis was extracted,and the minimum inhibitory concentration (MIC) of alcohol extract of S.baicalensis against drug-resistant E.coli was measured by agar dilution method,and the elimination rate was obtained by drug-resistance elimination test and photocopying method.Then,the drug-resistant changes of E.coli before and after drug-resistance elimination were detected by broth dilution and disk diffusion,the changes of bacterial morphology were observed by scanning electron microscopy (SEM),and the changes of relative expression of drug-resistance related genes were detected by Real-time quantitative PCR.【Result】 MIC of alcohol extract of S.baicalensis against apramycin-resistant E.coli was 41.25 mg/mL.The elimination rate of alcohol extract of S.baicalensis against apramycin-resistant E.coli was 11.7%-17.0%.The MIC of E.coli were reduced from 256 μg/mL to 16-32 μg/mL after alcohol extract of S.baicalensis treatment.Scanning electron microscope observation found that the surface of E.coli in alcohol extract of S.baicalensis treatment group was damaged and the morphology was abnormal. Real-time quantitative PCR detection showed that the mRNA relative expression of genes ompF,fimA,phnE and fhuF in E.coli was extremely significantly increased by alcohol extract of S.baicalensis (P<0.01).【Conclusion】 Alcohol extract of S.baicalensis could eliminate the drug-resistance of apramycin-resistant E.coli by upregulating the expression of ompF,fimA,phnE and fhuF genes,damaging the surface and improving the susceptibility of E.coli.

Key words: S.baicalensis; E.coli; drug-resistance elimination; apramycin

中图分类号:

S853.74

田二杰, 谷振, 王子皓, 刘方正, 刘克森, 张雪飞, 周变华. 黄芩对安普霉素耐药大肠杆菌的耐药消除研究[J]. 中国畜牧兽医, 2023, 50(3): 1279-1286.

TIAN Erjie, GU Zhen, WANG Zihao, LIU Fangzheng, LIU Kesen, ZHANG Xuefei, ZHOU Bianhua. Study on the Drug-resistance Elimination of Scutellaria baicalensis on Apramycin-resistant Escherichia coli[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1279-1286.

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