• 1.上海交通大學附屬第六人民醫(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，實驗結束后收集小鼠結腸組織，進行病理評分和形態(tài)觀察，ELISA法檢測結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技術對小鼠結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腸上皮細胞緊密連接蛋白進行定量和定位研究。結果與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構破壞; 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，阻止緊密連接超微結構破壞，促進緊密連接蛋白的表達(P lt;0.01)，降低結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。結論LP能通過促進腸上皮細胞間緊密連接蛋白的表達修復受損腸屏障功能，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. 中國普外基礎與臨床雜志, 2011, 18(12): 1242-1247. doi: 復制

版權信息： ?四川大學華西醫(yī)院華西期刊社《中國普外基礎與臨床雜志》版權所有，未經授權不得轉載、改編

1. Guarner F, Malagelada JR. Gut flora in health and disease [J]. Lancet, 2003, 361(9356): 512519.
2. Sartor RB. Microbial influences in inflammatory bowel diseases [J]. Gastroenterology, 2008, 134(2): 577594.
3. Shanahan F. Inflammatory bowel disease: immunodiagnostics, immunotherapeutics, and ecotherapeutics [J]. Gastroenterology, 2001, 120(3): 622635.
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. 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. 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. 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.