生物技术通报 ›› 2021, Vol. 37 ›› Issue (2): 103-110.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0743

• 研究报告 • 上一篇    下一篇

山姜素对鱼源耐药性嗜水气单胞菌体外抗菌作用的研究

陈杰豪1,2(), 缪玉佳2, 梁超2, 陶雨2, 欧阳萍2, 汪开毓2, 耿毅2, 石存斌1, 李宁求1()   

  1. 1.农业农村部渔用药物创制重点实验室 广东省水产动物免疫技术重点实验室 中国水产科学研究院珠江水产研究所,广州 510380
    2.四川农业大学动物医学院,成都 611130
  • 收稿日期:2020-06-17 出版日期:2021-02-26 发布日期:2021-02-26
  • 作者简介:陈杰豪,男,硕士研究生,研究方向:水生动物疾病学;E-mail: 530700885@qq.com
  • 基金资助:
    农业农村部渔用药物创制重点实验室开放基金课题(201702)

Study on the Antibacterial Mechanism of Alpinetin Against Fish-derived Drug-resistant Aeromonas hydrophila in vitro

CHEN Jie-hao1,2(), MIAO Yu-jia2, LIANG Chao2, TAO Yu2, OUYANG Ping2, WANG Kai-yu2, GENG Yi2, SHI Cun-bin1, LI Ning-qiu1()   

  1. 1. Key Laboratory of Fishery Drug Development,Ministry of Agriculture and Rural Affairs,Key Laboratory of Aquatic Animal Immune Technology,Guangdong Province,Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510380
    2. College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130
  • Received:2020-06-17 Published:2021-02-26 Online:2021-02-26

摘要:

旨为探究山姜素对耐药性嗜水气单胞菌的体外抑菌效果及其作用机制。通过测定山姜素对嗜水气单胞菌生长、细菌形态、电导率、乳酸脱氢酶(LDH)、蛋白质代谢和DNA的影响,研究山姜素对耐药性嗜水气单胞菌的抑菌机制。山姜素对5株鱼源耐药性嗜水气单胞菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)分别为128-256 μg/mL和512-1 024 μg/mL。与对照组相比,2MIC山姜素作用于嗜水气单胞菌CW株8 h、16 h后,菌体皱缩,细胞壁和细胞膜破损,细胞质丢失,内部空化;作用2 h后,菌悬液电导率、乳酸脱氢酶含量、DNA外渗量分别增长了3.97%、26.09%和10.50 μg/mL;作用2、4和8 h后,菌悬液核酸荧光强度和密度呈现随时间降低的趋势。山姜素对嗜水气单胞菌的体外抑菌机制为损伤菌体的细胞壁、增加细胞膜的通透性来抑制其生长繁殖。

关键词: 山姜素, 嗜水气单胞菌, 抑菌机制, 菌体形态

Abstract:

The aim of this study is to investigate the antibacterial effect and the mechanism of alpinetin against drug-resistant Aeromonas hydrophila in vitro. We investigated the mechanism of alpinetin against drug-resistant A. hydrophila by measuring the effects of alpinetin on A. hydrophila cell growth,cell morphology,electrical conductivity,lactate dehydrogenase(LDH),protein metabolism and DNA. The minimal inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of alpinetin against 5 strains of fish-derived A. hydrophila were 128-256 μg/mL and 512-1 024 μg/mL,respectively. Compared with the control group,after treating A. hydrophila with 2 MIC alpinetin for 8 or 16 h,the bacteria showed shrinkage,disrupted cell wall and membrane,loss of cytoplasm,and interior cavitation. After treating for 2 h,bacteria suspension’s electrical conductivity,LDH activity content and DNA extravasation increased by 3.97%,26.09% and 10.50 μg/mL,respectively. After treating for 2,4 and 8 h,nucleic acid fluorescence intensity and density in the bacterial suspension showed a time-dependent decreasing tendency. The in vitro antibacterial mechanism of alpinetin against A. hydrophila is to inhibit their growth and reproduction mainly via destroying cell walls of bacteria and increasing the permeability of the cell membrane.

Key words: alpinetin, Aeromonas hydrophila, antibacterial mechanism, cell morphology