摘要
以迟缓爱德华菌（E.tarda）胞外产物（ECP）为研究对象,通过饱和硫酸铵法从E.tarda培养上清中纯化得到其ECP,利用动物模型分析其致病性,并通过细胞试验证实了该产物对巨噬细胞功能影响,进一步测定其ECP的酶活性,并利用LC-MSMS方法分析E.tarda ECP蛋白成分。结果表明：迟E.tarda ECP对斑马鱼具有致病性,其具有淀粉酶、脂肪酶、卵磷脂酶活性和溶血活性可能在这一过程中发挥关键作用。除此之外,E.tarda ECP还能够显著增强巨噬细胞吞噬细菌活性的能力。LC-MSMS分析鉴定所提纯的ECP含有110种蛋白成分,涉及多种酶类、细胞结构蛋白和功能性蛋白。本研究为揭示E.tarda致病机理,研制新型疫苗奠定了基础。
Extracellular products（ECP）play an important role in pathogenesis of pathogenic bacteri- a. To investigate the roles of ECP in pathogenesis of Edwardsiella tarda, in this study, ECP was extracted and purified from E. tarda cultures at phase of logarithm. Then,the zebra fish model and Raw264.7 cells model were performed for assessing the the pathogenic and phagocytotic function of ECP,followed further enzyme activity tests and LC-MSMS. The data showed that the ECP of E. tarda was pathogenic to zebra fish, and could significantly increases the phagocytic activity of macrophage without bacterial species specificity, and the amylase, lipase, lecithin enzyme activity and hemolysis activity of ECP may play a ctitical role in this process. 110 proteins,covering a vari- ety of protein enzymes,cell componemts and functional protein, were identified to be components of ECP by LC-MSMS analysis. All these work provides guidance and foundation for exploring pathogenesis of E. tarda and development of subunit vaccine.
作者
张志强
李永慧
杨楠
苏硕青
羊扬
康元环
王洪彬
吴同垒
史秋梅
ZHANG Zhi-qiang1 , LI Yong-hui2 , YANG Nan1 , SU Shuo-qing1 , YA WANG Hong-bin1 , WU Tong-lei1. , SHI Qiu-mei1(1. Hebei Normal University of Science ＆ Technology, Qinhuangdao, Hebei 066600, China ; 2. The Second Hospital of Qinhuangdao , Qin- huangdao, Hebei 066600, China ; 3. Yangzhou University, Yangzhou , Jiangsu 225009, China ; 4. College of Anita al Science and Technology, J ilin Agriculture University, Changchun 130118, Chi-n)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2018年第11期2141-2146,共6页
Chinese Journal of Veterinary Science
基金
河北科技师范学院博士启动基金资助项目(2015YB002)
河北省高等学校科学技术研究资助项目(ZD2016067)
国家级星火计划资助项目(2015GA620002)
河北省科技厅科技支撑计划资助项目(14826613D)
