枸杞槲皮素对金黄色葡萄球菌产β-内酰胺酶的抑制作用

doi:10.16431/j.cnki.1671-7236.2024.01.039

摘要/Abstract

摘要： 【目的】探讨枸杞槲皮素对金黄色葡萄球菌产生的β-内酰胺酶的抑制作用机制，为临床研发β-内酰胺酶抑制剂提供基础依据。【方法】通过青霉素抑菌圈边缘试验筛选产β-内酰胺酶阳性菌株，通过药敏纸片法检测阳性菌株在32、64、128 μg/mL枸杞槲皮素作用下对青霉素的敏感性变化；借助多功能酶标仪检测在32、64、128、256 μg/mL枸杞槲皮素处理下青霉素含量的变化，并以此来评估枸杞槲皮素对β-内酰胺酶合成及在64、128、256 μg/mL枸杞槲皮素作用下对β-内酰胺酶活性的影响。通过AutoDockTools 1.5.6软件对枸杞槲皮素与BlaZ蛋白进行分子对接，采用Discovery Studio 2019 Client软件对分子对接结果进行可视化分析。【结果】从36株金黄色葡萄球菌中共筛选出23株产β-内酰胺酶阳性菌株。药敏纸片结果表明，随着枸杞槲皮素浓度的升高，阳性菌株对青霉素的敏感性增加。多功能酶标仪检测结果表明，枸杞槲皮素可以抑制β-内酰胺酶的合成，并显著抑制β-内酰胺酶的活性（P<0.05）。分子对接结果表明，枸杞槲皮素通过与BlaZ蛋白Ω-loop区域内的保守残基Asn161以氢键结合，影响BlaZ蛋白与β-内酰胺类抗生素的结合。【结论】枸杞槲皮素可以减少β-内酰胺酶的合成，还可通过降低β-内酰胺酶的活性增强青霉素的抑菌作用，具有作为β-内酰胺酶抑制剂的潜在活性，为后续开发新型β-内酰胺酶抑制剂提供了参考依据。

关键词: 金黄色葡萄球菌; 枸杞槲皮素; β-内酰胺酶; β-内酰胺酶抑制剂; 分子对接

Abstract: 【Objective】 This study was aimed to investigate the inhibitory mechanism of Lycium barbarum quercetin on β-lactamase produced by Staphylococcus aureus, so as to provide a basic basis for the clinical development of β-lactamase inhibitors.【Method】 β-lactamase positive strains were screened by penicillin inhibition zone edge test, and the sensitivity of positive strains to penicillin was detected by drug sensitivity paper method under the action of Lycium barbarum quercetin at concentrations of 32, 64 and 128 μg/mL.The changes of penicillin content under the treatment of Lycium barbarum quercetin at concentrations of 32, 64, 128 and 256 μg/mL were detected by multifunctional microplate reader, so as to evaluate the effect of Lycium barbarum quercetin on β-lactamase synthesis and β-lactamase activity under the treatment of Lycium barbarum quercetin at concentrations of 64, 128 and 256 μg/mL.The molecular docking of Lycium barbarum quercetin and BlaZ protein was performed by AutoDockTools 1.5.6 software, and the results of molecular docking were visualized by Discovery Studio 2019 Client software.【Result】 A total of 23 strains producing β-lactamase positive strains were screened from 36 strains of Staphylococcus aureus.The results of drug sensitivity paper showed that the sensitivity of positive strains to penicillin increased with the increase of Lycium barbarum quercetin concentration.The results of multifunctional microplate reader showed that Lycium barbarum quercetin could inhibit the synthesis of β-lactamase and significantly inhibit the activity of β-lactamase(P<0.05).Molecular docking results showed that Lycium barbarum quercetin affected the binding of BlaZ protein to β-lactam antibiotics by hydrogen bonding with the conserved residue Asn161 in the Ω-loop region of BlaZ protein.【Conclusion】 Lycium barbarum quercetin could reduce the synthesis of β-lactamase, and also enhance the antibacterial effect of penicillin by reducing the activity of β-lactamase, which had potential activity as a β-lactamase inhibitor.The results provided a reference for the subsequent development of new β-lactamase inhibitors.

Key words: Staphylococcus aureus; Lycium barbarum quercetin; β-lactamase; β-lactamase inhibitor; molecular docking

中图分类号:

S815

罗小凤, 刘盼盼, 陈晓慧, 张贝贝, 马彦军, 王桂琴. 枸杞槲皮素对金黄色葡萄球菌产β-内酰胺酶的抑制作用[J]. 中国畜牧兽医, 2024, 51(1): 382-391.

LUO Xiaofeng, LIU Panpan, CHEN Xiaohui, ZHANG Beibei, MA Yanjun, WANG Guiqin. Inhibitory Effect of Lycium barbarum Quercetin on β-lactamase from Staphylococcus aureus[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(1): 382-391.

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