四黄止痢复方醇提物(CSME)抗禽大肠杆菌的作用机制  

Antibacterial mechanism of methanol extract from Compound Sihuangzhili(CSME) against avian Escherichia coli

作　　者：孙嘉一 蒋红 马巍 刘全[2] 董国英 刘悦 曹艳子 周铁忠 王宏军 SUN Jia-yi;JIANG Hong;MA Wei;LIU Quan;DONG Guo-ying;LIU Yue;CAO Yan-zi;ZHOU Tie-zhong;WANG Hong-jun(Institute of Animal Husbandry and Veterinary Medicine ,Jinzhou Medical University , Jinzhou , Liaoning 121001, China;Animal Health Monitorand Warning Center of Liaoning Province , Shenyang 110001, China;College of Global Change and Earth System Science , Beijing Normal University , Beijing 100875 China;Dalian Sanyi Animal Medicine Co. 9LTD., Dalian ,Liaoning 116036, China)

机构地区：[1]锦州医科大学畜牧兽医学院,辽宁锦州121001 [2]辽宁省动物卫生监测预警中心,辽宁沈阳110001 [3]北京师范大学全球变化与地球系统科学研究院,北京100875 [4]大连三仪动物药品有限公司,辽宁大连116036

出　　处：《中国兽医学报》2019年第3期493-496,506,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31201951;31601914);辽宁省高等学校优秀人才基金资助项目(LJQ2013087)

摘　　要：为了探讨四黄止痢复方醇提物(CSME)对禽大肠杆菌的抗菌机制。采用微孔-平板法检测其对禽大肠杆菌的最小抑菌浓度(MIC);用吸光光度法检测其对细菌生长曲线、细胞膜通透性和能量代谢活力的影响;荧光定量PCR法检测其对禽大肠杆菌4种主要毒力因子(tsh、iss、ibeA和fimC)表达的影响。结果显示,CSME对禽大肠杆菌(CVCC251)的MIC为125 g/L,1/2MIC就能显著抑制细菌生长,破坏细菌细胞膜结构,使核酸、蛋白质外泄,并抑制了琥珀酸脱氢酶(SDH)和苹果酸脱氢酶(MDH)的活性;1/8MIC能显著抑制了禽大肠杆菌tsh、iss和fimC毒力因子基因的表达(P<0.01)。结果表明,CSME可以通过破坏细菌细胞膜结构、造成细菌能量代谢紊乱、抑制毒力基因表达发挥抗菌活性。In order to study the antibacterial mechanism of CSME against avian Escherichia coli(E.coli),CSME was extracted with methanol.Minimum inhibitory concentration(MIC) and the effect of the extract against the E.coli growth curve was explored by micropore-plate and absorptiometry.Effect of the extract against cell membrane permeability and energy metabolism of avian E.coli was determined by spectrophotometry.The influence of the extract against virulence factors gene expression of tsh,iss,ibeA and fimC in avian E.coli,were evaluated by fluorescence quantitative PCR.The results showed that MIC of the extract against avian E.coli was 125 g/L and subinhibitory concentration could inhibit the growth of E.coli in the early stages.The extract could increase bacterial cell membrane permeability and lead to bacterial nucleic acid and protein macromolecular material overflow.The extract could inhibit the activity of SDH and MDH and the gene expression of tsh,iss and fimC in avian E.coli.CSME methanol extract could destroy cell membrane structure,causing energy metabolism disorder and inhibiting virulence gene expression.

关 键 词：四黄止痢复方 提取物 抗菌机制 禽大肠杆菌 

分 类 号：S859.3[农业科学—临床兽医学]