• 1.上海交通大學(xué)附屬第六人民醫(yī)院普外科(上海 200233);;
  • 2.上海交大昂立醫(yī)藥研究院(上海 200233);

目的研究益生菌植物乳桿菌(LP)對炎癥狀態(tài)下腸屏障的修復(fù)機理。方法以白介素-10基因敲除(IL-10－/－)小鼠為炎性腸病模型，采用簡單隨機法(隨機表)將空白對照(WT)小鼠和IL-10－/－小鼠分成4組: WT組、WT+LP組、IL-10－/－組和IL-10－/－+LP組，每組10只。分別灌飼Ringer液或LP溶液，持續(xù)4周。每周記錄小鼠腸炎的臨床表現(xiàn)，實驗結(jié)束后收集小鼠結(jié)腸組織，進行病理評分和形態(tài)觀察，ELISA法檢測結(jié)腸組織促炎細胞因子腫瘤壞死因子-α(TNF-α)和干擾素-γ(IFN-γ)濃度，利用Ussing Chamber技術(shù)對小鼠結(jié)腸通透性及電生理變化進行分析，并用Western blot和免疫熒光分析法對結(jié)腸上皮細胞緊密連接蛋白進行定量和定位研究。結(jié)果與WT組相比，IL-10－/－組小鼠腹瀉、直腸脫垂和體重喪失較明顯(P lt;0.01); TNF-α和IFN-γ濃度明顯升高(P lt;0.01); 病理表現(xiàn)為大量炎性細胞浸潤，偶見透壁性潰瘍和隱窩膿腫形成; 超微電鏡提示緊密連接結(jié)構(gòu)破壞; 緊密連接蛋白(ZO-1、occludin、claudin-1)表達明顯降低(P lt;0.01)且伴有異常分布; 結(jié)腸上皮細胞甘露醇通透性升高(P lt;0.001)，跨膜電阻值降低(P lt;0.001)。LP干預(yù)4周后，與IL-10－/－組相比，LP能顯著改善腸道炎癥的臨床和病理表現(xiàn)，阻止緊密連接超微結(jié)構(gòu)破壞，促進緊密連接蛋白的表達(P lt;0.01)，降低結(jié)腸上皮細胞甘露醇通透性(P lt;0.001)，升高跨膜電阻值(P lt;0.001)，降低了TNF-α和IFN-γ的濃度(P lt;0.01)，減輕了腸道炎癥。結(jié)論LP能通過促進腸上皮細胞間緊密連接蛋白的表達修復(fù)受損腸屏障功能，從而減輕腸道炎癥。

引用本文： 陳紅旗,楊俊,周玉坤,張旻,杭曉敏,秦環(huán)龍. 益生菌對炎性腸病小鼠腸屏障功能的影響. 中國普外基礎(chǔ)與臨床雜志, 2011, 18(12): 1242-1247. doi: 復(fù)制

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7. Madsen K, Cornish A, Soper P, et al. Probiotic bacteria enhance murine and human intestinal epithelial barrier function [J]. Gastroenterology, 2001, 121(3): 580591.
8. Yu Y, Sitaraman S, Gewirtz AT. Intestinal epithelial cell regulation of mucosal inflammation [J]. Immunol Res, 2004, 29(13): 5568.
9. Munkholm P, Langholz E, Hollander D, et al. Intestinal permeability in patients with Crohn’s disease and ulcerative colitis and their first degree relatives [J]. Gut, 1994, 35(1): 6872.
10. Peeters M, Geypens B, Claus D, et al. Clustering of increased small intestinal permeability in families with Crohn’s disease[J]. Gastroenterology， 1997, 113(3): 802807.
11. Prasad S, Mingrino R, Kaukinen K, et al. Inflammatory processes have differential effects on claudins 2, 3 and 4 in colonic epithelial cells [J]. Lab Invest, 2005, 85(9): 11391162.
12. DarfeuilleMichaud A, Boudeau J, Bulois P, et al. High prevalence of adherentinvasive Escherichia coli associated with ileal mucosa in Crohn’s disease [J]. Gastroenterology, 2004, 127(2): 412421.
13. Martin HM, Campbell BJ, Hart CA, et al. Enhanced Escherichia coli adherence and invasion in Crohn’s disease and colon cancer [J]. Gastroenterology, 2004, 127(1): 8093.
14. Vanderpool C, Yan F, Polk DB. Mechanisms of probiotic action: Implications for therapeutic applications in inflammatory bowel diseases [J]. Inflamm Bowel Dis, 2008, 14(11): 15851596.
15. JohnsonHenry KC, Donato KA, ShenTu G, et al. Lactobacillus rhamnosus strain GG prevents enterohemorrhagic Escherichia coli O157: H7induced changes in epithelial barrier function [J]. Infect Immun, 2008, 76(4): 13401348.
16. RestaLenert S, Barrett KE. Probiotics and commensals reverse TNFalpha and IFNgammainduced dysfunction in human intestinal epithelial cells [J]. Gastroenterology, 2006, 130(3): 731746.
17. RestaLenert S, Barrett KE. Live probiotics protect intestinal epithelial cells from the effects of infection with enteroinvasive Escherichia coli (EIEC) [J]. Gut, 2003, 52(7): 988997.
18. Qin HL, Zhang ZW, Hang XM, et al. L. plantarum prevents enteroinvasive Escherichia coliinduced tight junction proteins changes in intestinal epithelial cells [J]. BMC Microbiol, 2009, 9: 6371.
  1. 1. Guarner F, Malagelada JR. Gut flora in health and disease [J]. Lancet, 2003, 361(9356): 512519.
  2. 2. Sartor RB. Microbial influences in inflammatory bowel diseases [J]. Gastroenterology, 2008, 134(2): 577594.
  3. 3. Shanahan F. Inflammatory bowel disease: immunodiagnostics, immunotherapeutics, and ecotherapeutics [J]. Gastroenterology, 2001, 120(3): 622635.
  4. 4. Hedin C, Whelan K, Lindsay JO. Evidence for the use of probiotics and prebiotics in inflammatory bowel disease: a review of clinical trials [J]. Proc Nutr Soc, 2007, 66(3): 307315.
  5. 5. Berg DJ, Davidson N, Kühn R, et al. Enterocolitis and colon cancer in interleukin10deficient mice are associated with aberrant cytokine production and CD4(+) TH1like responses [J]. J Clin Invest, 1996, 98(4): 10101020.
  6. 6. Zeissig S, Bürgel N, Günzel D, et al. Changes in expression and distribution of claudin 2, 5 and 8 lead to discontinuous tight junctions and barrier dysfunction in active Crohn’s disease [J]. Gut, 2007, 56(1): 6172.
  7. 7. Madsen K, Cornish A, Soper P, et al. Probiotic bacteria enhance murine and human intestinal epithelial barrier function [J]. Gastroenterology, 2001, 121(3): 580591.
  8. 8. Yu Y, Sitaraman S, Gewirtz AT. Intestinal epithelial cell regulation of mucosal inflammation [J]. Immunol Res, 2004, 29(13): 5568.
  9. 9. Munkholm P, Langholz E, Hollander D, et al. Intestinal permeability in patients with Crohn’s disease and ulcerative colitis and their first degree relatives [J]. Gut, 1994, 35(1): 6872.
  10. 10. Peeters M, Geypens B, Claus D, et al. Clustering of increased small intestinal permeability in families with Crohn’s disease[J]. Gastroenterology， 1997, 113(3): 802807.
  11. 11. Prasad S, Mingrino R, Kaukinen K, et al. Inflammatory processes have differential effects on claudins 2, 3 and 4 in colonic epithelial cells [J]. Lab Invest, 2005, 85(9): 11391162.
  12. 12. DarfeuilleMichaud A, Boudeau J, Bulois P, et al. High prevalence of adherentinvasive Escherichia coli associated with ileal mucosa in Crohn’s disease [J]. Gastroenterology, 2004, 127(2): 412421.
  13. 13. Martin HM, Campbell BJ, Hart CA, et al. Enhanced Escherichia coli adherence and invasion in Crohn’s disease and colon cancer [J]. Gastroenterology, 2004, 127(1): 8093.
  14. 14. Vanderpool C, Yan F, Polk DB. Mechanisms of probiotic action: Implications for therapeutic applications in inflammatory bowel diseases [J]. Inflamm Bowel Dis, 2008, 14(11): 15851596.
  15. 15. JohnsonHenry KC, Donato KA, ShenTu G, et al. Lactobacillus rhamnosus strain GG prevents enterohemorrhagic Escherichia coli O157: H7induced changes in epithelial barrier function [J]. Infect Immun, 2008, 76(4): 13401348.
  16. 16. RestaLenert S, Barrett KE. Probiotics and commensals reverse TNFalpha and IFNgammainduced dysfunction in human intestinal epithelial cells [J]. Gastroenterology, 2006, 130(3): 731746.
  17. 17. RestaLenert S, Barrett KE. Live probiotics protect intestinal epithelial cells from the effects of infection with enteroinvasive Escherichia coli (EIEC) [J]. Gut, 2003, 52(7): 988997.
  18. 18. Qin HL, Zhang ZW, Hang XM, et al. L. plantarum prevents enteroinvasive Escherichia coliinduced tight junction proteins changes in intestinal epithelial cells [J]. BMC Microbiol, 2009, 9: 6371.