酪酸梭菌-糯米复合发酵液对鼠伤寒沙门菌的抑制作用及其对鼠伤寒沙门菌感染小鼠肠道分泌型免疫球蛋白A的影响  

Inhibitory effect of Clostridium butyricum-glutinous rice compound fermentation liquid on Salmonella typhimurium and its effect on secretory immunoglobulin A in intestine of mice infected with Salmonella typhimurium

作　　者：姜林娟 郑志[1,2] 朱绍辉 朴丙熙[2,4] JIANG Linjuan;ZHENG Zhi;ZHU Shaohui;PARK Byounghee(Department of Toxicology,School of Public Health,Xinxiang Medical University,Xinxiang 453003,Henan Province,China;Hanling Medical Technology Research Institute Co.,Nanjing 211100,Jiangsu Province,China;Department of General Surgery,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China;Healing Co.,Seoul 04393,Republic of Korea)

机构地区：[1]新乡医学院公共卫生学院卫生毒理学教研室,河南新乡453003 [2]韩陵医药研究院有限公司,江苏南京211100 [3]新乡医学院第一附属医院普通外科,河南卫辉453100 [4]喜灵生物科技有限公司,韩国首尔04393

出　　处：《新乡医学院学报》2022年第2期113-121,共9页Journal of Xinxiang Medical University

基　　金：新乡医学院海外归国博士启动基金项目(编号:505098);河南省自然科学基金项目(编号:162300410221);河南省高等学校重点科研项目计划(编号:22B320007)。

摘　　要：目的探讨酪酸梭菌-糯米复合发酵液对鼠伤寒沙门菌的抑制作用及其对小鼠肠道分泌型免疫球蛋白A(sIgA)的影响。方法将酪酸梭菌(1.5×10^(10) CFU·L^(-1))和体积分数5%的无菌糯米粉放入到100 mL胰蛋白胨大豆肉汤,于37℃条件下培养48 h,配制酪酸梭菌-糯米复合发酵液,采用显微镜平板菌落计数方法检测酪酸梭菌-糯米复合发酵液和酪酸梭菌悬液中酪酸梭菌,琼脂打孔体外抑菌圈法检测酪酸梭菌-糯米复合发酵液及其稀释液(1:2、1:4、1:8、1:16)对鼠伤寒沙门菌的抑制作用,显微镜平板菌落计数方法检测酪酸梭菌悬液、无细胞发酵液、酪酸梭菌-糯米复合发酵液、糯米发酵液、磷酸盐缓冲液(PBS)、四环素对鼠伤寒沙门菌的抑制作用。应用200μL鼠伤寒沙门菌悬液(1×10^(12) CFU·L^(-1))对60只雄性6周龄ICR小鼠进行灌胃,每日1次,连续灌胃7 d构建鼠伤寒沙门菌感染小鼠模型,将鼠伤寒沙门菌感染小鼠随机分为糯米发酵液组、酪酸梭菌悬液组、无细胞发酵液组、酪酸梭菌-糯米复合发酵液组、PBS阴性对照组、四环素阳性对照组,每组10只,分别给予酪酸梭菌悬液、无细胞发酵液、酪酸梭菌-糯米复合发酵液、糯米发酵液、PBS、体积分数0.2%四环素各200μL灌胃,每日1次,连续灌胃21 d,应用显微镜平板菌落计数方法检测小鼠体内鼠伤寒沙门菌菌株数,酶联免疫吸附测定法检测肠道内sIgA表达水平,监测小鼠食物摄取量和体质量,计算体质量增加率。结果培养24 h后,酪酸梭菌-糯米复合发酵液中酪酸梭菌菌落数显著多于酪酸梭菌悬液(P<0.05)。酪酸梭菌-糯米复合发酵液对鼠伤寒沙门菌的抑制作用呈剂量-效应关系(P<0.05)。体外抑菌试验结果显示,酪酸梭菌悬液、无细胞发酵液、酪酸梭菌-糯米复合发酵液、四环素均对鼠伤寒沙门菌具有抑制作用,PBS和糯米发酵液对鼠伤寒沙门菌无抑菌作用。体内抑菌试�Objective To investigate the inhibitory effect of Clostridium butyricum(C.butyricum)-glutinous rice compound fermentation liquid on Salmonella typhimurium(S.typhimurium)and its effect on secretory immunoglobulin A(sIgA)in intestine of mice infected with S.typhimurium.Methods The C.butyricum(1.5×10^(10) CFUμL^(-1))and volume fraction of 5% sterile glutinous rice powder were put into 100 mL tryptone soybean broth and cultured for 48 h at 37℃ to prepare C.butyricum-glutinous rice compound fermentation liquid.The colony number of C.butyricum in the C.butyricum-glutinous rice compound fermentation liquid and C.butyricum suspension was detected by using plate counting method.The inhibitory effect of C.butyricum-glutinous rice compound fermentation liquid and its diluents(1:2,1:4,1:8,1:6)on S.typhimurium was detected by using agar diffusion method.The inhibitory effects of the C.butyricum suspension,acellular fermentation broth,C.butyricum-glutinous rice compound fermentation liquid,glutinous rice fermentation broth,phosphate buffered solution(PBS)and tetracycline on S.typhimurium were detected by the plate counting method.The 60 male 6-week-old ICR mice were orally administrated with 200μL S.typhimurium suspension(1×1012 CFUμL^(-1))to establish S.typhimurium infection mouse model,once a day for 7 days,and they were randomly divided into C.butyricum suspension group,acellular fermentation broth group,C.butyricum-glutinous rice compound fermentation liquid group,glutinous rice fermentation broth group,PBS negative control group and tetracycline positive control group,with 10 mice in each group,and the mice were respectively treated with 200μL of C.butyricum suspension,200μL of acellular fermentation broth,200μL of C.butyricum-glutinous rice fermentation liquid,200μL of glutinous rice fermentation broth,200μL of PBS and 200μL of volume fraction of 0.2% tetracycline,once a day for 21 days,the number of S.typhimurium in intestine of mice was detected by plate counting method,and the level of sIgA in intestine o

关 键 词：酪酸梭菌 鼠伤寒沙门菌 分泌型免疫球蛋白A 抑菌作用 

分 类 号：R117[医药卫生—公共卫生与预防医学] R-33