蜂毒肽与依布硒啉对金黄色葡萄球菌的协同作用与机制研究  

Study on the Synergistic Effect of Ebselen and Melittin Against Staphylococcus aureus and the Possible Mechanism

作　　者：刘彬彬 周靖轩 蔡轶 张赶 王君 杜幼芹[1] Liu Binbin;Zhou Jingxuan;Cai Yietal(Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy,Institute of Infection and Inflammatory Damage,Medicine College of Three Gorges University,Yichang 443002,China)

机构地区：[1]三峡大学肿瘤微环境与免疫治疗湖北省重点实验室,三峡大学医学院感染与炎症损伤研究所,宜昌443002

出　　处：《华中科技大学学报（医学版）》2023年第1期36-42,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81903105);湖北省卫生健康委联合基金重点项目(No.WJ2019H527);宜昌市医疗卫生科研项目(No.A19-301-50)。

摘　　要：目的研究蜂毒肽(Melittin,Mel)与依布硒啉(Ebselen,EbSe)对金黄色葡萄球菌(Staphylococcus aureus,S.aureus)的协同抗菌活性和机制。方法96微孔板实验结合分光光度法(visible spectrophotometry,UV-vis)检测蜂毒肽-依布硒啉的抗金黄色葡萄球菌作用;电子显微镜(electron microscopy,EM)观察蜂毒肽-依布硒啉处理后金黄色葡萄球菌的细胞形态。通过5,5′-二硫代双(2-硝基苯甲酸)[5,5′-Dithiobis-(2-nitrobenzoic acid),DTNB]获得电子后显色的能力,观察蜂毒肽-依布硒啉对细菌硫氧蛋白还原酶(thioredoxin reductase,TrxR)活性的影响。通过流式细胞术(flow cytometry,FCM)检测蜂毒肽-依布硒啉对细菌胞内活性氧(reactive oxygen species,ROS)产量的影响;使用活性氧清除剂二硫苏糖醇(dithiothreitol,DTT)复测上述指标。结果蜂毒肽与依布硒啉具有协同杀菌作用,可协同抑制金黄色葡萄球菌TrxR,显著上调金黄色葡萄球菌胞内活性氧产量。DTT挽救实验证明蜂毒肽-依布硒啉对细菌造成损伤是通过活性氧途径实现。结论蜂毒肽可通过协同依布硒啉靶向金黄色葡萄球菌TrxR,上调胞内活性氧的大量产生,介导细菌死亡。Objective To study the synergistic antibacterial effect of Melittin(Mel)and Ebselen(EbSe)against Staphylococcus aureus(S.aureus).Methods Visible spectrophotometry(UV-vis)and 96-microwell plate assay were performed to detect the antibacterial effect of Mel-EbSe against S.aureus;electron microscope(EM)was used to observe the morphological change of S.aureus after Mel-EbSe treatment;5,5′-Dithiobis-(2-nitrobenzoic acid)(DTNB)assay was used to detect the effect of Mel-EbSe on TrxR activity;flow cytometry was used to detect the effect of Mel-EbSe reactive oxygen species(ROS)production in bacteria;ROS scavenger dithiothreitol(DTT)was used to retest the above indicators.Results Mel-EbSe had a significant synergistic inhibitory effect against S.aureus.Mel-EbSe could effectively inhibit the activity of TrxR,and significantly induced the production of ROS in S.aureus.DTT rescue test showed that Mel-EbSe caused damage to bacteria through reactive oxygen species.Conclusion Mel-EbSe has an antibacterial effect on S.aureus through upregulation of ROS by co-targeting TrxR.

关 键 词：蜂毒肽 依布硒啉 金黄色葡萄球菌 活性氧 

分 类 号：R378.11[医药卫生—病原生物学]