湖北医药学院学报2024,Vol.43Issue(5) :463-469.DOI:10.13819/j.issn.2096-708X.2024.05.001

原薯蓣皂苷体外抗产NDM-1耐碳青霉烯Raoultella ornithinolytica B1645-1B的机制研究

Mechanism of Protodioscin Against Carbapenem-resistant Raoultella Ornithinolytica B1645-1B Carrying NDM-1 in Vitro

余春芳 严廷苇 妥娟 彭紫晶 张思涵 胡景秀 何浩 张涵肃 孟丽雪
湖北医药学院学报2024,Vol.43Issue(5) :463-469.DOI:10.13819/j.issn.2096-708X.2024.05.001

原薯蓣皂苷体外抗产NDM-1耐碳青霉烯Raoultella ornithinolytica B1645-1B的机制研究

Mechanism of Protodioscin Against Carbapenem-resistant Raoultella Ornithinolytica B1645-1B Carrying NDM-1 in Vitro

余春芳 1严廷苇 2妥娟 3彭紫晶 4张思涵 4胡景秀 4何浩 4张涵肃 4孟丽雪5
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作者信息

  • 1. 湖北医药学院基础医学院;武当特色中药研究湖北省重点实验室
  • 2. 湖北医药学院药护学院
  • 3. 湖北医药学院第一临床学院
  • 4. 湖北医药学院第二临床学院,湖北 十堰 442000
  • 5. 湖北医药学院基础医学院
  • 折叠

摘要

目的:研究原薯蓣皂苷(protodioscin,PD)对产碳青霉烯酶NDM-1 解鸟氨酸乌拉尔菌(carbapenem-re-sistant Raoultella ornithinolytica)B1645-1B的体外抑菌作用及逆转耐药性机制研究.方法:采用微量肉汤稀释法测定PD对B1645-1B的最低抑菌浓度(minimum inhibitory concentration,MIC).应用棋盘法测定中药单体PD联合亚胺培南(Imipenem,IMP)对B1645-1B的MIC,计算抑菌浓度指数(fractional inhibitory concentration,FIC).采用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测PD作用菌株B1645-1B后NDM-1 mRNA合成.利用MOE(Molecular Operating Environment)软件预测PD与NDM-1 蛋白分子对接.通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE)检测PD作用纯化后菌株B1645-1B的NDM-1 蛋白表达.结果:PD对B1645-1B 的 MIC 为10 000 μg/mL.PD联合IMP对B1645-1B的 FIC为0.75,出现协同作用.PD作用后菌株B1645-1B 的NDM-1 mRNA合成抑制和酶活性降低.结论:PD可能是一种很有前途的新型有效的协同抗菌剂及NDM-1 酶抑制剂.

Abstract

Objective To investigate the antibacterial effect of protodioscin(PD)against carbapenem-resistant Raoultella ornithinolytica B1645-1B carrying NDM-1 in vitro and its mechanism.Methods A broth microdilution method was used to determine the minimum inhibitory concentration(MIC)of PD against strain B1645-1B.The MIC of PD combined with imi-penem(IMP)against the strain was determined by the checkerboard method,and the fractional inhibitory concentration(FIC)index was calculated.The mRNA expression of NDM-1 in strain B1645-1B after treatment with PD was detected by quantitative real-time PCR(qRT-PCR).The Molecular Operating Environment(MOE)was used to predict the molecular docking between PD and NDM-1 protein.The protein expression of NDM-1 after treatment with PD and purification was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).Results The MIC of PD against strain B1645-1B was 10 000 μg/mL,and PD combined with IMP had an FIC of 0.75,indicating synergy.After treatment with PD,the mRNA synthesis and enzyme activity of NDM-1 were inhibited.Conclusion PD was a promising novel and effective synergistic antimicrobial agent and NDM-1 enzyme inhibitor.

关键词

原薯蓣皂苷/耐碳青霉烯解鸟氨酸乌拉尔菌/NDM-1/亚胺培南/体外抑菌实验

Key words

Protodioscin/Raoultella ornithinolytica/NDM-1/Imipenem/Antibacterial experiment in vitro

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基金项目

湖北省教育厅自然科学基金(2023AFB270)

湖北省大学生创新创业训练计划项目(S202213249002)

十堰市科技局项目(22Y23)

十堰市科技局项目(22Y21)

出版年

2024
湖北医药学院学报
湖北医药学院

湖北医药学院学报

影响因子:0.504
ISSN:1006-9674
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