[1]姜林娟,郑　志,朱绍辉,等.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[J].新乡医学院学报,2022,39(2):113-121.[doi:10.7683/xxyxyxb.2022.02.003]
　JIANG Linjuan,ZHENG Zhi,ZHU Shaohui,et al.Inhibitory effect ofClostridium butyricum-glutinous rice compound fermentation liquid on S. typhimurium and its effect on secretory immunoglobulin A in intestine of mice infected withS. typhimurium[J].Journal of Xinxiang Medical University,2022,39(2):113-121.[doi:10.7683/xxyxyxb.2022.02.003]
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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
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
39
期数:
2022年2
页码:
113-121
栏目:
基础研究
出版日期:
2022-02-05

文章信息/Info

Title:
Inhibitory effect ofClostridium butyricum-glutinous rice compound fermentation liquid on S. typhimurium and its effect on secretory immunoglobulin A in intestine of mice infected withS. typhimurium
作者:
姜林娟1; 郑　志1; 2; 朱绍辉3; 朴丙熙2; 4
（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）
Author(s):
JIANG Linjuan1; ZHENG Zhi1; 3; ZHU Shaohui2; PARK Byounghee3; 4
(1.Department of Toxicology,School of Public Health,Xinxiang Medical University,Xinxiang 453003,Henan Province,China;2.Hanling Medical Technology Research Institute Co.,Nanjing 211100,Jiangsu Province,China;3.Department of General Surgery,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China;4.Healing Co.,Seoul 04393,Republic of Korea)
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
Keywords:
Clostridium butyricum; Salmonella typhimurium; secretory immunoglobulin A; antimicrobial effect
分类号:
R117，R-33
DOI:
10.7683/xxyxyxb.2022.02.003
文献标志码:
A
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（1.5×1010 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200 μL鼠伤寒沙门菌悬液（1×1012 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200 μL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。体内抑菌试验结果显示，PBS阴性对照组和糯米发酵液组第0、7、14、21、28天小鼠体内鼠伤寒沙门菌呈增高趋势（P<0.05）；四环素阳性对照组、酪酸梭菌悬液组、无细胞发酵液组、酪酸梭菌-糯米复合发酵液组小鼠体内鼠伤寒沙门菌菌落数均出现先增高后降低趋势（P<0.05），且酪酸梭菌-糯米复合发酵液组小鼠体内鼠伤寒沙门菌菌落数第28天与第0天比较差异无统计学意义（P>0.05）。第0、7、14天，6组小鼠体内鼠伤寒沙门菌菌落数比较差异无统计学意义（P>0.05）；第21天，四环素阳性对照组和酪酸梭菌-糯米复合发酵液组小鼠体内鼠伤寒沙门菌菌落数显著少于PBS阴性对照组、糯米发酵液组、酪酸梭菌悬液组和无细胞发酵液组（P<0.05）。第28天，四环素阳性对照组、酪酸梭菌悬液组、无细胞发酵液组、酪酸梭菌-糯米复合发酵液组小鼠体内鼠伤寒沙门菌菌落数显著少于PBS阴性对照组和糯米发酵液组，四环素阳性对照组、酪酸梭菌-糯米复合发酵液组小鼠体内鼠伤寒沙门菌菌落数显著少于酪酸梭菌悬液组和无细胞发酵液组，酪酸梭菌-糯米复合发酵液组小鼠体内鼠伤寒沙门菌菌落数显著少于四环素阳性对照组（P<0.05）。酪酸梭菌-糯米复合发酵液组小鼠肠道内sIgA水平显著高于四环素阳性对照组、酪酸梭菌悬液组、无细胞发酵液组、糯米发酵液组和PBS阴性对照组，四环素阳性对照组小鼠肠道内sIgA水平显著高于酪酸梭菌悬液组、无细胞发酵液组、糯米发酵液组和PBS阴性对照组，酪酸梭菌悬液组小鼠肠道内sIgA水平显著高于无细胞发酵液组、糯米发酵液组和PBS阴性对照组，无细胞发酵液组小鼠肠道内sIgA水平显著高于糯米发酵液组和PBS阴性对照组（P<0.05）。四环素阳性对照组和糯米发酵液组小鼠的食物摄取量高于酪酸梭菌悬液组、PBS阴性对照组、无细胞发酵液组、酪酸梭菌-糯米复合发酵液组（P<0.05）；酪酸梭菌悬液组和酪酸梭菌-糯米复合发酵液组小鼠的体质量增加量高于PBS阴性对照组、四环素阳性对照组、无细胞发酵液组、糯米发酵液组（P<0.05），四环素阳性对照组、无细胞发酵液组、糯米发酵液组小鼠的体质量增加量高于PBS阴性对照组（P<0.05）；四环素阳性对照组、糯米发酵液组、酪酸梭菌悬液组、无细胞发酵液组、酪酸梭菌-糯米复合发酵液组小鼠的体质量增加率高于PBS阴性对照组，酪酸梭菌悬液组、无细胞发酵液组、酪酸梭菌-糯米复合发酵液组小鼠的体质量增加率高于四环素阳性对照组、糯米发酵液组（P<0.05）。结论　酪酸梭菌-糯米复合发酵液可显著抑制鼠伤寒沙门菌生长，提高机体免疫力，这为多重耐药鼠伤寒沙门菌感染膳食干预策略的制定提供了新的思路和依据。
Abstract:
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 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 200 μL S.typhimurium suspension (1×1012 CFU μL-1) to establish S.typhimurium infection mouse model,once a day for 7 days, the number of S.typhimurium in intestine of mice was detected by plate counting method,and the level of sIgA in intestine of mice were determined by using enzyme linked immunosorbent assay.In addition,the food intake and body mass of mice were monitored.Results　The colony number of C.butyricum grown in the C.butyricum-glutinous rice compound fermentation liquid group was significantly higher than that in the C.butyricum suspension group after 24 hours of culture (P<0.05).The inhibitory effect of C.butyricum-glutinous rice compound fermentation liquid on S.typhimurium showed a dose-response relationship(P<0.05).The C.butyricum suspension,sterile fermentation broth,C.butyricum-glutinous rice compound fermentation liquid and tetracycline had inhibitory effects on S.typhimurium,while PBS and glutinous rice fermentation broth had no inhibitory effects on S.typhimurium. The colony number of S.typhimurium in intestine of mice in the PBS negative control group and the glutinous rice compound fermentation broth group was increased (P<0.05);the colony number of S.typhimurium in intestine of mice in the tetracycline positive control group,C.butyricum suspension group,acellular fermentation broth group and C.butyricum-glutinous rice compound fermentation liquid group were beginning to increase and then decrease (P<0.05),and there was no significant difference in the colony number of S.typhimurium in intestine of mice in the C.butyricum-glutinous rice compound fermentation liquid group between day 28 and day 0 (P>0.05).On day 0,7 and 14,there was no significant difference in the colony number of S.typhimurium in intestine of mice among the six groups (P>0.05);On the 21st day,the colony number of S.typhimurium in intestine of mice in the tetracycline positive control group and C.butyricum-glutinous rice compound fermentation liquid group was significantly less than that in the PBS negative control group,glutinous rice compound fermentation liquid group,C.butyricum suspension group and acellular fermentation liquid group (P<0.05).On the 28th day,the colony number of S.typhimurium in intestine of mice in the tetracycline positive control group,C.butyricum suspension group,acellular fermentation liquid group and C.butyricum-glutinous rice compound fermentation liquid group was significantly less than that in the PBS negative control group and glutinous rice fermentation liquid group,the colony number of S.typhimurium in intestine of mice in the tetracycline positive control group and C.butyricum-glutinous rice compound fermentation liquid was significantly less than that in the C.butyricum suspension group and acellular fermentation liquid group,the colony number of S.typhimurium in intestine of mice in the C.butyricum-glutinous rice compound fermentation liquid was significantly less than that in the tetracycline positive control group (P<0.05).The level of sIgA in intestine of mice in the C.butyricum-glutinous rice compound fermentation liquid group was significantly higher than that in the tetracycline positive control group,C.butyricum suspension group,acellular fermentation broth group,glutinous rice fermentation broth group and PBS negative control group (P<0.05);the level of sIgA in intestine of mice in the tetracycline positive control group was significantly higher than that in the C.butyricum suspension group,acellular fermentation broth group,glutinous rice fermentation broth group and PBS negative control group (P<0.05);the level of sIgA in intestine of mice in the C.butyricum suspension group was significantly higher than that in the acellular fermentation broth group,glutinous rice fermentation broth group and PBS negative control group(P<0.05);the level of sIgA in intestine of mice in the acellular fermentation broth group was significantly higher than that in the glutinous rice compound fermentation broth group and PBS negative control group (P<0.05).The increase of body mass of mice in the C.butyricum suspension group and C.butyricum-glutinous rice compound fermentation liquid group was significantly higher than that in the PBS negative control group,tetracycline positive control group,acellular fermentation liquid group and glutinous rice compound fermentation liquid group (P<0.05).The increase of body mass of mice in the tetracycline positive control group,acellular fermentation liquid group and glutinous rice compound fermentation liquid group was significantly higher than that in the PBS negative control group (P<0.05).The rate of body mass increase of mice in the tetracycline positive control group,glutinous rice fermentation broth group,C.butyricum suspension group,acellular fermentation broth group and C.butyricum-glutinous rice compound fermentation liquid group were higher than that in the PBS negative control group.The rate of body mass increase of mice in the C.butyricum suspension group,acellular fermentation broth group and C.butyricum-glutinous rice fermentation liquid group were higher than that in the tetracycline positive control group and glutinous rice fermentation broth group (P<0.05).Conclusion　C.butyricum-glutinous rice compound fermentation liquid can significantly inhibit S.typhimurium infection and improve immunity,which provides novel ideas and scientific basis for the formulation of dietary intervention plan for multi-drug resistant S.typhimurium infection.

参考文献/References:

［1］　SUN H,WAN Y,DU P,et al.The epidemiology of monophasic Salmonella typhimurium［J］.Foodborne Pathog Dis,2020,17(2):87-97.
［2］　SHEN H,CHEN H,OUY,et al.Prevalence,serotypes,and antimicrobial resistance of Salmonella isolates from patients with diarrhea in Shenzhen,China［J］.BMC Microbiol,2020,20(1):197.
［3］　张建群,罗学辉,袁士杰.余姚市2007-2018年感染性腹泻病原谱监测分析［J］.中国公共卫生管理,2020,36(6):868-871.
ZHANG J Q,LUO X H,YUAN S J.Surveillance and analysis of pathogenic spectrum of infectious diarrhea in Yuyao city from 2007-2018［J］.Chin J Public Healh Manag,2020,36(6):868-871.
［4］　李永瑞,李艳.河南省临床鼠伤寒沙门氏菌的耐药性分析［J］.河南科技大学学报,2018,36(3):218-221.
LI Y R,LI Y.Antibiotic resistance of Salmonella enterica serovar typhimurium in human infections in Henan［J］.J Henan Sci Technol Univ,2018,36(3):218-221.
［5］　辛洪杰,白杨.酪酸梭菌在胃肠道疾病中的研究与应用进展［J］.现代消化及介入诊疗,2020,25(5):677-680.
XIN H J,BAI Y.Progress in research and application of Clostridium butyricum in gastrointestinal disease［J］.Modern Digest Intervent,2020,25(5):677-680.
［6］　江舒曼,赵寒冰,贾林,等.酪酸梭菌对急性坏死性胰腺炎并发腹腔内高压大鼠肠道屏障功能及菌群的影响［J］.中华胰腺病杂志,2019,19(3):181-184.
JIANG S M,ZHAO H B,JIA L,et al.Effects of Clostridium butyricum on the function of intestinal mucosal barrier and flora in acute necrotizing pancreatitis rats with intra-abdominal hypertension［J］.Chin J Pancreatol,2019,19(3):181-184.
［7］　SHIN D S,EOM Y B.Antimicrobial and antibiofilm activities of Clostridium butyricum supernatant against Acinetobacter baumannii［J］.Arch Microbiol,2020,202(5):1059-1068.
［8］　刘岩红,袁凤云,蒋立会.酪酸梭菌活菌胶囊对行早期肠内营养支持的感染性休克患者胃肠道症状改善作用观察［J］.中国微生态学杂志,2021,33(2):209-213.
LIU Y H,YUAN F Y,JIANG L H.Effect of Clostridium butyricum capsule on gastrointestinal symptoms in patients with septic shock undergoing early enteral nutrition support［J］.China J Microecol,2021,33(2):209-213.
［9］　刘鑫,马中岭,张长存.复方茵陈糖浆联合益生菌与间歇光疗治疗新生儿母乳性黄疸疗效观察［J］.新乡医学院学报,2020,37(2):181-184.
LIU X,MA Z L,ZHANG C C.Curative effect of compound yinchen syrup combined with probiotics and intermittent phototherapy in the treatment of neonatal breast milk jaundice［J］.J Xinxiang Med Univ,2020,37(2):181-184.
［10］　TOMITA Y,IKEDA T,SAKATAS,et al.Association of probiotic Clostridium butyricum therapy with survival and response to immune checkpoint blockade in patients with lung cancer［J］.Cancer Immunol Res,2020,8(10):1236-1242.
［11］　DENKOVA R,YANAKIEVA V,DENKOVA Z,et al.In vitro inhibitory activity of bifidobacterium and lactobacillus strains against Candida albicans［J］.Bulg J Vet Med,2013,16(3):186-197.
［12］　高智群,林菁,朱炜春,等.儿童鼠伤寒沙门菌感染54例临床分析［J］.当代医学,2020,26(16):147-149.
GAO Z Q,LIN J,ZHU W C,et al.Clinical analysis of 54 children infected with Salmonella typhimurium［J］.Contemp Med,2020,26(16):147-149.
［13］　赵夜,林菁,赖宗乐,等.小儿复杂型鼠伤寒沙门菌感染治疗特点及药敏结果分析［J］.中国医学创新,2020,17(15):121-125.
ZHAO Y,LIN J,LAI Z L.et al.Treatment characteristics and drug sensitivity analysis of complex Salmonella typhimurium infection in children［J］.Med Innovat China,2020,17(15):121-125.
［14］　石国露.儿童非伤寒沙门氏菌感染临床特征及菌血症危险因素分析［D］.重庆:重庆医科大学,2020.
SHI G L.Clinical manifestation and risk factors for bacteremia of nontyphoidal Salmonslla infections in children［D］.Chongqing:Chongqing Medical University,2020.
［15］　CARROLL L M,GABALLA A,GULDIMANN C,et al.Identification of novel mobilized colistin resistance gene mcr-9 in a multidrug-resistant,colistin-susceptible salmonella enterica serotype typhimurium isolate［J］.mBio,2019,10(3):e00853-19.
［16］　TYSON G H,LI C,HARRISON L B,et al.Amultidrug-resistant Salmonella in fantisclone is spreading and recombining in the United States［J］.Microb Drug Resist,2021,27(6):792-799.
［17］　KARP B E,LEEPER M M,CHEN J C,et al.Multidrug-resistant Salmonellaserotype anatum in travelers and seafood from Asia,United States［J］.Emerg Infect Dis,2020,26(5):1030-1033.
［18］　王玉环.抗菌药物在儿童消化系统疾病中的使用策略［J］.世界临床药物,2020,41(12):941-946.
WANG Y H.Application strategy of antibacterial agents in pediatric gastroenterological disease［J］.World Clin Drugs,2020,41(12):941-946.
［19］　CHEN H,SHI T,LU Y,et al.Antimicrobial resistance,genetic diversity and virulence genes of Salmonella typhimurium isolated in infant with acute diarrhea in Fuzhou,China,2015-2017［J］.Clin Lab,2019,65(5)779-787.
［20］　ZHOU Y,XU W,HONG K,et al.Therapeutic effects of probiotic Clostridium butyricum WZ001 on bacterial vaginosis in mice［J］.J Appl Microbiol,2019,127(2):565-575.
［21］　张树波,崔云龙,吴顺娥,等.酪酸梭菌的抑菌作用研究［J］.中国新药杂志,2002,11(4):322-324.
ZHANG S B,CUI Y L,WU S E,et al.Inhibitory effect of Clostridium butyricum(CB) on bacteria［J］.Chin J New Drugs,2002,11(4):322-324.
［22］　马依晨,孙月宏,杨慧.还原型谷胱甘肽联合酪酸梭菌二联活菌散对新生儿黄疸胆红素水平及T细胞亚群指标的影响［J］.临床医学研究与实践,2021,6(13):81-83.
MA Y C,SUN Y H,YANG H.Effects of reduced glutathione combined with combined Clostridium butyricumand Bifidobacterium powders on bilirubin level and T cell subsets indexes in neonatal jaundice［J］.Clin Res Pract,2021,6(13):81-83.
［23］　GUO Q,GOLDENBERG J Z,HUMPHREY C,et al.Probiotics for the prevention of pediatric antibiotic-associated diarrhea［J］.Cochrane Database Syst Rev,2019,4(4):CD004827.
［24］　DOUMATEY A P,ADEYEMO A,ZHOU J,et al.Gutmicrobiome profiles are associated with type 2 diabetes in Urban Africans［J］.Front Cell Infect Microbiol,2020,10:63.
［25］　LIU M,XIE W,WAN X,et al.Clostridium butyricum protects intestinal barrier function via upregulation of tight junction proteins and activation of the Akt/mTOR signaling pathway in a mouse model of dextran sodium sulfate-induced colitis［J］.Exp Ther Med,2020,20(5):10.
［26］　于玲,刘志敏,曾海英,等.不同大米营养价值分析［J］.现代食品,2020,12(24):183-186.
YU L,LIU Z M,ZENG H Y,et al.Analysis of nutritional value of different rice［J］.Modern Food,2020,12(24):183-186.

更新日期/Last Update: 2022-02-05