甘草总黄酮对艰难梭菌的体内外抗菌作用研究  

In Vitro and in Vivo Antibacterial Activities of Total Flavonoids from Glycyrrhiza uralensis against Clostridium difficile

作　　者：孙剑峰[1] 任丽蓉 黄博 刘希望[2] SUN Jian-feng;REN Li-rong;HUANG Bo;LIU Xi-wang(Gansu Province Animal Disease Prevention and Control Center,Lanzhou,Gansu,730000,China;Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS/Key Laboratory of New Animal Drug Discovery,Ministry of Agriculture and Rural Affairs/Key Laboratory of New Animal Drug Project of Gansu Province,Lanzhou,Gansu,730050,China)

机构地区：[1]甘肃省动物疫病预防控制中心,甘肃兰州730000 [2]中国农业科学院兰州畜牧与兽药研究所/农业农村部兽用药物创制重点实验室/甘肃省新兽药工程重点实验室,甘肃兰州730050

出　　处：《动物医学进展》2024年第9期61-66,共6页Progress In Veterinary Medicine

基　　金：甘肃省自然科学基金项目(22JR5RA042);中国农业科学院科技创新工程(25-LZIHPS-02)。

摘　　要：考察甘草总黄酮对艰难梭菌的抑菌活性及其抗菌机制,为艰难梭菌感染(CDI)的防治和甘草的高效利用提供依据。用微量肉汤稀释法、计数法测定了甘草总黄酮对艰难梭菌的最小抑菌浓度(MIC)、最小杀菌浓度(MBC)、生长曲线和杀菌曲线。通过扫描电镜观察、碘化丙啶(PI)摄取试验、胞外蛋白和ATP含量的测定,检测了甘草总黄酮对细菌细胞膜完整性的影响。建立了小鼠CDI模型,以体重变化和存活率为指标,评价了甘草总黄酮的体内抗菌活性。结果表明,甘草总黄酮对艰难梭菌的MIC和MBC分别为12.5μg/mL和25μg/mL,具有快速杀菌的特征。甘草总黄酮可显著破坏细菌细胞膜,导致ATP和蛋白等胞内物质的泄露,使细菌形态发生显著改变,进而导致菌体死亡。甘草总黄酮可显著缓解CDI小鼠的体重下降,7 d存活率为100%,疗效与万古霉素相当。甘草总黄酮具有显著的体内外抗艰难梭菌活性,破坏细菌细胞膜的完整性是其可能的抗菌机制。The in vitro and in vivo antibacterial activities of total flavonoids from Glycyrrhiza uralensis(TFGU)against Clostridium difficile and the possible mode of action were investigated,which can provide basis for the prevention and treatment of CDI and the efficient utilization of licorice.Micro-broth dilution method and bacterial counting method were applied to determine the minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC).The growth curve and killing curve of TFGU on C.difficile were also evaluated.The membrane integrity was investigated by scanning electron microscope,propidium iodide(PI)uptake assay,determination of extracellular ATP and protein.Finally,CDI model was successfully established on mice and in vivo efficacy of TFGU was also evaluated.The results showed that MIC and MBC of TFGU against C.difficile were 12.5 and 25μg/mL,respectively.TFGU showed a rapid bactericidal manner.Bacterial membrane was significantly disrupted by TFGU,which lead to the leakage of ATP and protein,morphological change and finally death of bacteria.The body weight decrease of mice was significantly alleviated by TFGU and the 7-day survival rate was 100%,which was equal to vancomycin.In conclusion,TFGU showed significant in vitro and in vivo antibacterial activity against C.difficile and disruption of bacterial membrane maybe the possible mechanism of action.

关 键 词：艰难梭菌 甘草总黄酮 抗菌作用 细胞膜 

分 类 号：S853.7[农业科学—临床兽医学]