FHA结构域蛋白TagH对霍乱弧菌肠道定植能力及致病力的影响  

Effects of the FHA domain protein TagH on intestinal colonization and pathogenicity of Vibrio cholerae

作　　者：贾城壹 范婵[3] 卢琴 王小素 闵迅 黄健 Jia Chengyi;Fan Chan;Lu Qin;Wang Xiaosu;Min Xun;Huang Jian(Department of Laboratory Medicine,the Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563099,China;School of Laboratory Medicine,Zunyi Medical University,Zunyi Guizhou 563099,China;Department of Laboratory Medicine,Suining Central Hospital,Suining Sichuan 629099,China)

机构地区：[1]遵义医科大学附属医院医学检验科,贵州遵义563099 [2]遵义医科大学检验医学院,贵州遵义563099 [3]遂宁市中心医院检验科,四川遂宁629099

出　　处：《遵义医科大学学报》2023年第5期431-437,共7页Journal of Zunyi Medical University

基　　金：国家自然科学青年基金资助项目(NO:31800130);贵州省科技厅基础研究计划项目[NO:黔科合基础-ZK(2021)一般470]。

摘　　要：目的探究FHA结构域蛋白TagH对非O1/非O139群霍乱弧菌肠道定植能力和致病力的影响,并探讨这种影响是否依赖于溶血素HlyA和蛋白酶PrtV的表达。方法通过生长曲线实验测定各菌株的生长速率。取CD1乳鼠和CD1小鼠各分为PBS阴性对照组、WT菌株感染组、ΔtagH菌株感染组、ΔtagHΔhlyA菌株感染组和ΔtagHΔprtV菌株感染组,通过乳鼠肠道定植实验分析各菌株的肠道定植能力;通过小鼠灌胃感染实验测定各菌株感染组小鼠外周血白细胞数量以及IL-6、IL-8和TNF-α水平;观察各菌株感染组小鼠回肠组织病理损伤及7 d的生存情况。结果细菌生长曲线实验发现,各菌株的生长速度基本一致并无明显差异。乳鼠肠道定植实验发现,与WT菌株相比,ΔtagH菌株的肠道定植能力增强(P<0.01),ΔtagHΔhlyA菌株的肠道定植能力较ΔtagH菌株进一步增强(P<0.05),ΔtagHΔprtV菌株的肠道定植能力弱于ΔtagH菌株(P<0.05)。小鼠灌胃感染实验发现,与WT菌株相比,ΔtagH菌株的白细胞数升高(P<0.05)且肠组织损伤更严重,ΔtagHΔhlyA菌株的白细胞数较ΔtagH菌株明显下降(P<0.05)且几乎没有肠组织损伤,ΔtagHΔprtV菌株感染现象与ΔtagH菌株相似。WT菌株感染组、ΔtagH菌株感染组、ΔtagHΔhlyA菌株感染组和ΔtagHΔprtV菌株感染组的小鼠生存率分别为62.5%、75%、100%和75%。结论TagH抑制霍乱弧菌的肠道定植能力和致病力,其调控作用主要依赖于霍乱弧菌溶血素HlyA,而蛋白酶PrtV的作用不明显。Objective To investigate the effect of the FHA domain protein TagH on intestinal colonization ability and pathogenicity of non-O1/non-O139 Vibrio cholerae,and to explore whether this effect is dependent on the expression of hemolysis HlyA and protease PrtV.Methods The growth rate of each strain was measured by growth curve experiment.CD1 suckling mice and CD1 mice were respectively divided into PBS negative control group,WT strain infection group,ΔtagH strain infection group,ΔtagHΔhlyA strain infection group andΔtagHΔprtV strain infection group.The intestinal colonization ability of each strain was analyzed by intestinal colonization experiment of suckling mice.The number of peripheral white blood cells and the levels of IL-6,IL-8 and TNF-αwere determined by intragastric infection in mice.Pathological injury of ileal tissue and 7-day survival of mice in each strain infection group were observed.Results The experiment of bacterial growth curve showed that the growth rate of each strain was basically the same and there was no obvious difference.The intestinal colonization experiment of suckling mice showed that compared with WT strain,the intestinal colonization ability ofΔtagH strain was enhanced(P<0.01),the intestinal colonization ability ofΔtagHΔhlyA strain was further enhanced than that ofΔtagH strain(P<0.05),and the intestinal colonization ability ofΔtagHΔprtV strain was weaker than that ofΔtagH strain(P<0.05).Intragastric infection in mice showed that compared with WT strain,ΔtagH strain had higher white blood cell count(P<0.05)and more severe intestinal tissue damage,ΔtagHΔhlyA strain had significantly lower white blood cell count thanΔtagH strain(P<0.05)and almost no intestinal tissue damage,and the infection phenomenon ofΔtagHΔprtV strain was similar to that ofΔtagH strain.The survival rates of WT group,ΔtagH group,ΔtagHΔhlyA group andΔtagHΔprtV group were 62.5%,75%,100%and 75%,respectively.Conclusion TagH inhibits the intestinal colonization ability and pathogenicity of Vibrio cho

关 键 词：FHA结构域蛋白TagH 非O1/非O139群霍乱弧菌 溶血素HlyA 蛋白酶PrtV 

分 类 号：R378.3[医药卫生—病原生物学]