摘要
为了探讨天麻蛋白对金黄色葡萄球菌(金葡菌)的抑制作用,本研究以德江天麻为原料,提取天麻蛋白,检测天麻蛋白在不同浓度和不同时间条件下对金葡菌抑菌率的影响.利用实时荧光定量PCR检测金葡菌的毒力基因的表达变化,用试剂盒检测天麻蛋白对金葡菌细胞外碱性磷酸酶含量的影响和总肠毒素表达量的影响,用细菌计数法检测天麻蛋白对感染金葡菌小鼠死亡保护率和肺部细菌数量的影响.结果显示,天麻蛋白分子量分布于25~35 kDa,其对金葡菌的抑菌率呈现出剂量依赖性,天麻蛋白的抑菌率随着与金葡菌作用时间的延长而增加;天麻蛋白可抑制金葡菌毒力基因sea,agrA,hla及总肠毒素的表达,又可增加金葡菌细胞外碱性磷酸酶含量,并且均具有浓度依赖性;天麻蛋白降低了金葡菌肺炎小鼠死亡率,减少了小鼠肺脏中金葡菌的定植数量.以上结果说明,天麻蛋白在宿主体内外均可抑制金葡菌的生长.
In order to investigate the inhibitory effect of Gastrodin elata protein on Staphylococcus aureus(S.aureus),Gastrodin elata of Dejiang was used as a raw material to extract Gastrodin elata protein,and effects of Gastrodia elata protein on the antibacterial rate of S.aureus under different concentrations and different time conditions were detected in this study.The expression changes of virulence genes of S.aureus was detected by real-time fluorescence quantitative PCR.Effects of Gastrodia elata protein on the extracellular alkaline phosphatase content and total enterotoxin expression of S.aureus were detected by kits.Effects of Gastrodia elata protein on the death protection rate and the number of lung bacteria in mice infected with S.aureus were detected by bacterial counting method.The results showed that the molecular weight of Gastrodia elata protein is in the range of 25~35 kDa.The inhibitory rate of Gastrodia elata protein to S.aureus is dose-dependent and is gradually enhanced with the prolongation of the interaction time with S.aureus.Expression levels of virulence genes sea,agrA,hla,and total enterotoxin of S.aureus are inhibited by S.aureus,and the extracellular alkaline phosphatase content of S.aureus is enhanced by Gastrodia elata in a concentration-dependent manner.The mortality of S.aureus pneumonia mice and the number of S.aureus colonization in the lungs of mice are decreased by Gastrodia elata protein.All results showed that the growth of S.aureus is inhibited by Gastrodia elata protein both in vitro and in vivo.
作者
印双红
张俊波
张孟琴
易萌
张红
YIN Shuang-hong;ZHANG Jun-bo;ZHANG Meng-qin;YIN Meng;ZHANG Hong(College of One Health,Tongren University,Tongren 554300,Guizhou,China;Guizhou Provincial Key Laboratory for Biodiversity Conservation and Utilization in the Fanjing Mountain Region,Tongren University,Tongren 554300,Guizhou,China;College of Agroforestry Engineering and Planning,Tongren University,Tongren 554300,Guizhou,China;College of Materials and Chemical Engineering,Tongren 554300,Guizhou,China)
出处
《西北师范大学学报(自然科学版)》
CAS
北大核心
2021年第6期88-93,共6页
Journal of Northwest Normal University(Natural Science)
基金
贵州省教育厅青年科技人才成长项目(黔教合KY字[2017]312)
贵州省科技支撑项目(黔科合支撑[2019]2348)
贵州省高层次创新人才培养项目(2018-(2016)-022)
贵州省重点实验室项目(黔科合平台人才[2020]2003号)
铜仁市科技支撑计划项目(铜市科研[2020]128号)
铜仁学院高端人才创新团队项目(CXTD[2020-19])。
关键词
德江天麻蛋白
金葡菌
毒力基因
小鼠
抑制作用
Gastrodia elata protein of Dejiang
Staphylococcus aureus
virulence genes
mice
inhibition