益生菌抵御大鼠急性腹泻的效果及其对肠道黏膜屏障的影响

庞敏; 卢庆萍; 朱丽媛; 夏冰; 张宏福

doi:10.3969/j.issn.1006-267x.2016.05.022

动物营养学报 >

2016 , Vol. 28 >Issue 5: 1462 - 1470

DOI: https://doi.org/10.3969/j.issn.1006-267x.2016.05.022

饲料营养 Feed Science and Technology

益生菌抵御大鼠急性腹泻的效果及其对肠道黏膜屏障的影响

庞敏 ,
卢庆萍 ,
朱丽媛 ,
夏冰 ,
张宏福

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中国农业科学院北京畜牧兽医研究所, 动物营养学国家重点实验室, 北京 100193

收稿日期: 2015-12-07

网络出版日期: 2016-05-24

基金资助

国家科技支撑项目(2012BAD39B01);中国农业科学院创新团队(ASTIP-IAS07)

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Effects of Probiotics to Against Acute Diarrhea and on Intestinal Mucosal Barrier of Rats

PANG Min ,
LU Qingping ,
ZHU Liyuan ,
XIA Bing ,
ZHANG Hongfu

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State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing 100193, China

Received date: 2015-12-07

Online published: 2016-05-24

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摘要

本试验旨在研究酪酸梭菌或唾液乳酸菌对预防急性腹泻、保护肠道黏膜屏障的作用效果。选取清洁级21日龄断奶Wistar大鼠[平均体重(57.45±6.23) g]48只,随机分在4个组:正常对照组(C1组)、模型对照组(C2组)、酪酸梭菌组(T1组)、唾液乳酸菌组(T2组)。每个组6个重复,每个重复2只大鼠。试验期共9 d,C1、C2组经口灌服生理盐水, T1、T2组分别灌服酪酸梭菌和唾液乳酸菌。第8天开始除C1组外,其余各组灌服番泻叶制剂致泻,记录各组大鼠腹泻情况。试验结束后处死所有大鼠并采集血液及肠道样品。结果显示:1)灌服番泻叶后各组大鼠均出现腹泻。C2组稀便率、腹泻指数、血清内毒素及D-乳酸含量极显著高于C1组(P<0.01);而与C2组相比,T1、T2组腹泻指数、血清内毒素及D-乳酸含量均极显著降低(P<0.01)。2)与C2组相比,T1、T2组隐窝深度显著降低(P<0.05),绒毛高度与隐窝深度的比值极显著升高(P<0.01),且T1组回肠闭锁小带蛋白1(ZO-1)、闭锁蛋白(occludin) mRNA的相对表达量分别极显著(P<0.01)和显著(P<0.05)升高。3)C2组回肠白细胞介素-10(IL-10)、白细胞介素-1β(IL-1β)mRNA的相对表达量分别极显著(P<0.01)和显著(P<0.05)高于C1组;而与C2组相比,T1组回肠IL-10 mRNA的相对表达量极显著降低(P<0.01)。以上结果说明,在本试验中,酪酸梭菌与唾液乳酸菌在一定程度上可以缓解由番泻叶引起的大鼠急性腹泻,降低肠道通透性,同时具有增强和保护肠道黏膜屏障的作用。

关键词： 酪酸梭菌; 唾液乳酸菌; 大鼠; 急性腹泻; 肠道黏膜屏障

本文引用格式

庞敏 , 卢庆萍 , 朱丽媛 , 夏冰 , 张宏福 . 益生菌抵御大鼠急性腹泻的效果及其对肠道黏膜屏障的影响[J]. 动物营养学报, 2016 , 28(5) : 1462 -1470 . DOI: 10.3969/j.issn.1006-267x.2016.05.022

Abstract

The purpose of this study was to evaluate the effects of Clostridium butyricum or Lactobacillus salivarius ZJ614 on prevention of acute diarrhea and protection of intestinal mucosal barrier function. Forty-eight 21-day-weanned Wistar rats with the average body weight of (57.45±6.23) g were selected and randomly divided into four groups:normal control group (C1 group), model control group (C2 group), Clostridium butyricum group (T1 group) and Lactobacillus salivarius ZJ614 group (T2 group). Each group had six replicates, and each replicate had two rats. This experiment lasted for 9 days. C1 and C2 groups were orally administered saline, and T1 and T2 groups were administered probiotics accordingly. On the 8th day, every group except C1 group was administered folium senna to cause acute diarrhea and then record diarrhea condition of rats. All rats were sacrificed, and blood and intestinal samples were collected at the end of experiment. The results showed as follows:1) all rats displayed diarrhea symptoms as long as administered folium senna. Loose stools rate, diarrhea index, serum endotoxin and D-lactic acid contents were significantly increased in C2 group compared with C1 group (P<0.01). The diarrhea index, serum endotoxin and D-lactic acid contents decreased significantly in T1 and T2 groups compared with C2 group (P<0.01). 2) Compared with C2 group, the crypt depth in T1 and T2 groups was significantly decreased (P<0.05), the ratio of villus height to crypt depth in T1 and T2 groups was significantly increased (P<0.01), and the relative expression levels of ileum zonula occludens-1(ZO-1) (P<0.01) and occludin mRNA (P<0.05) in T1 group were significantly improved. 3) C2 group significantly up-regulated the relative expression levels of ileum interleukin-10(IL-10) (P<0.01)and interleukin-1β (IL-1β) mRNA (P<0.05) compared with C1 group, and T1 group down-regulated the relative expression level of IL-10 mRNA compared with C2 group (P<0.01). In summary, Clostridium butyricum or Lactobacillus salivarius ZJ614 can alleviate the actue diarrhea induced by senna on a certain degree, and also decrease the intestinal permeability, and enhance intestinal mucosal barrier.

Key words： Clostridium butyricum; Lactobacillus salivarius ZJ614; rats; actue diarrhea; intestinal mucosal barrier

参考文献

[1] CAMILLERI M,NULLENS S,NELSEN T.Enteroendocrine and neuronal mechanisms in pathophysiology of acute infectious diarrhea[J].Digestive Diseases and Sciences,2012,57(1):19-27.

[2] 徐立新.嗜酸乳杆菌和亚油酸治疗抗生素引起大鼠腹泻的研究[D].硕士学位论文.泸州:泸州医学院,2011:4-5.
[3] 曾娟.肠易激综合征肠黏膜屏障功能的研究[D].博士学位论文.济南:山东大学,2008:10-12.
[4] GAREAU M G,SHERMAN P M,WALKER W A.Probiotics and the gut microbiota in intestinal health and disease[J].Nature Reviews Gastroenterology & Hepatology,2010,7(9):503-514.

[5] SUN Y J,CAO H J,SONG D D,et al.Probiotics can alleviate cardiopulmonary bypass-induced intestinal mucosa damage in rats[J].Digestive Diseases and Sciences,2013,58(6):1528-1536.

[6] GUARINO A,GUANDALINI S,LO VECCHIO A.Probiotics for prevention and treatment of diarrhea[J].Journal of Clinical Gastroenterology,2015,49(Suppl.1):S37-S45.
[7] OHLAND C L,MACNAUGHTON W K.Probiotic bacteria and intestinal epithelial barrier function[J].American Journal of Physiology:Gastrointestinal and Liver Physiology,2010,298(6):G807-G819.
[8] LING Z X,LIU X,CHENG Y W,et al.Clostridium butyricum combined with Bifidobacterium infantis probiotic mixture restores fecal microbiota and attenuates systemic inflammation in mice with antibiotic-associated diarrhea[J].BioMed Research International,2015,2015:582048.
[9] 胡长霞.益生菌对轮状病毒性腹泻小鼠肠道屏障的影响及机制的探讨[D].硕士学位论文.武汉:华中科技大学,2013:5-6.
[10] TAKAHASHI M,TAGUCHI H,YAMAGUCHI H,et al.The effect of probiotic treatment with Clostridium butyricum on enterohemorrhagic Escherichia coli O157:H7 infection in mice[J].FEMS Immunology & Medical Microbiology,2004,41(3):219-226.

[11] BISSON J F,HIDALGO S,ROZAN P,et al.Preventive effects of different probiotic formulations on travelers' diarrhea model in Wistar rats:preventive effects of probiotics on TD[J].Digestive Diseases and Sciences,2010,55(4):911-919.

[12] PALOMAR M M,GALDEANO C M,PERDIGÓN G.Influence of a probiotic lactobacillus strain on the intestinal ecosystem in a stress model mouse[J].Brain,Behavior,and Immunity,2014,35:77-85.
[13] 李丹,吕妍,唐方.腹泻型肠易激综合征大鼠模型的建立[J].天津中医药,2009,26(3):240-242.
[14] 周干南,胡芝华,汪亚先,等.小鼠腹泻模型的制备与腹泻指数的应用[J].中草药,1994,25(4):195-196,199.
[15] 王新,韩月东,吴汉平,等.番泻叶提取物对小鼠胃肠运动及其组织形态学的影响[J].中国中西医结合杂志,2001,21(增刊):66-68.
[16] 兰梅,王新,吴汉平,等.番泻叶提取物对人肠上皮细胞生物学特性的影响[J].世界华人消化杂志,2001,9(5):555-559.
[17] SONG Z F,WU T X,CAI L S,et al.Effects of dietary supplementation with Clostridium butyricum on the growth performance and humoral immune response in Miichthys miiuy[J].Journal of Zhejiang University Science B,2006,7(7):596-602.

[18] 刘宗正,徐轲,王剑,等.酪酸菌对腹泻小鼠的治疗效果及其安全性[J].山东大学学报:医学版,2009,47(9):32-35.
[19] VENTOLA H,LEHTORANTA L,MADETOJA M,et al.Effects of the viability of Lactobacillus rhamnosus GG on rotavirus infection in neonatal rats[J].World Journal of Gastroenterology,2012,18(41):5925-5931.

[20] JOHNSTON S D,SMYE M,WATSON R P.Intestinal permeability tests in coeliac disease[J].Clinical Laboratory,2000,47(3/4):143-150.
[21] 李亚杰,赵献军.益生菌对肠道黏膜免疫的影响[J].动物医学进展,2006,27(7):38-41.
[22] LUYER M D,BUURMAN W A,HADFOUNE M,et al.Strain-specific effects of probiotics on gut barrier integrity following hemorrhagic shock[J].Infection and Immunity,2005,73(6):3686-3692.

[23] 刘虎传,张敏红,李素霞,等.益生菌制剂对早期断奶仔猪肠道pH、黏膜形态结构和挥发性脂肪酸含量的影响[J].动物营养学报,2012,24(7):1329-1335.
[24] BRANISTE V,JOUAULT A,GAULTIER E,et al.Impact of oral bisphenol A at reference doses on intestinal barrier function and sex differences after perinatal exposure in rats[J].Proceedings of the National Academy of Sciences of the United States of America,2010,107(1):448-453.

[25] EBNET K,AURRAND-LIONS M,KUHN A,et al.The junctional adhesion molecule (JAM) family members JAM-2 and JAM-3 associate with the cell polarity protein PAR-3:a possible role for JAMs in endothelial cell polarity[J].Journal of Cell Science,2003,116(19):3879-3891.

[26] TURNER J R.Molecular basis of epithelial barrier regulation:from basic mechanisms to clinical application[J].The American Journal of Pathology,2006,169(6):1901-1909.

[27] ENDO H,NIIOKA M,KOBAYASHI N,et al.Butyrate-producing probiotics reduce nonalcoholic fatty liver disease progression in rats:new insight into the probiotics for the gut-liver axis[J].PLoS One,2013,8(5):e63388.
[28] MANTIS N J,ROL N,CORTHÉSY B.Secretory IgA's complex roles in immunity and mucosal homeostasis in the gut[J].Mucosal Immunology,2011,4(6):603-611.

[29] SHOUVAL D S,OUAHED J,BISWAS A,et al.Interleukin 10 receptor signaling:master regulator of intestinal mucosal homeostasis in mice and humans[J].Advances in Immunology,2014,122:177-210.
[30] SHOUVAL D S,BISWAS A,GOETTEL J A,et al.Interleukin-10 receptor signaling in innate immune cells regulates mucosal immune tolerance and anti-inflammatory macrophage function[J].Immunity,2014,40(5):706-719.

[31] MORAN C J,WALTERS T D,GUO C H,et al.IL-10R polymorphisms are associated with very-early-onset ulcerative colitis[J].Inflammatory Bowel Diseases,2012,doi:10.1002/ibd.22974.
[32] 任大勇.益生乳酸杆菌的黏附及免疫调节作用研究[D].博士学位论文.长春:吉林大学,2013:4-6.
[33] COCCIA M,HARRISON O J,SCHIERING C,et al.IL-1β mediates chronic intestinal in-flammation by promoting the accumulation of IL-17A secreting innate lymphoid cells and CD4⁺ Th17 cells[J].The Journal of Experimental Medicine,2012,209(9):1595-1609.

[34] NEURATH M F.Cytokines in inflammatory bowel disease[J].Nature Reviews Immunology,2014,14(5):329-342.

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