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肠内营养联合益生菌对腹部感染大鼠肠道菌群及屏障功能的影响

Influences of enteral nutrition combined with probiotics on gut microflora and barrier function of rats with abdominal infection.

作者信息

Shen Tong-Yi, Qin Huan-Long, Gao Zhi-Guang, Fan Xiao-Bing, Hang Xiao-Ming, Jiang Yan-Qun

机构信息

Department of Surgery, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai 200233, China.

出版信息

World J Gastroenterol. 2006 Jul 21;12(27):4352-8. doi: 10.3748/wjg.v12.i27.4352.

Abstract

AIM

To investigate the influences of enteral, parenteral nutrition and probiotics delivered by gut on intestinal microecology, epithelial tight junctions, immune and barrier function of rats with abdominal infection.

METHODS

Rat abdominal infection models established with cecal ligation and perforation method, were divided into three groups: parenteral nutrition (PN group, n = 7), PN+enteral nutrition (EN group, n = 7) and PN + EN + probiotics (probiotics group, n = 7) via the needle jejunostomy and neck vein for five days. The total nutritional supplement of the three groups was isonitrogenic and isocaloric. Probiotics was delivered by jejunostomy 10 mL/d (1 x 10(8) cfu/mL). The rats were killed on the sixth day. The feces in the cecum were cultured for anaerobic bacterial growth and analyzed with bacterial group DNA fingerprint profile with random amplified polymorphic DNA. The transmembrane binding proteins (occludin) and IgA level in plasma cells of intestine epithelium in colon and terminal ileum were measured by an immunohistochemistry method. The ultrastructure of intestinal epithelial tight junctions in colon and small intestine was observed by electron-microscopy. Vena cava blood and the homogenated tissue of liver, lung and mesenteric lymph nodes were cultured to determine the bacterial translocations, and endotoxin in the blood from portal vein was detected.

RESULTS

(1) The amount of bacteria of gut species in EN group and probiotic group was higher than that in PN group. The DNA-profiles in EN group and probiotic group were similar to that of normal rats. The number of DNA-profiles in probiotics group was much more than that in PN group and EN group. Moreover, there were strange stripes in PN group. (2) The expression of occludin and IgA in the small and large intestine in EN group (2.309 +/- 0.336, 15.440 +/- 2.383) and probiotic group (2.938 +/- 0.515, 16.230 +/- 3.183) was improved as compared with PN group (1.207 +/- 0.587, P < 0.05, 11.189 +/- 2.108, P < 0.01). The expression of occludin in probiotic group (intestine: 2.93 +/- 0.515; cecum: 3.40 +/- 0.617) was higher than that in EN group (intestine: 2.309 +/- 0.336; cecum: 2.076 +/- 0.670; P < 0.05). The expression of IgA, especially in EN group (intestine: 15.440 +/- 2.383) and probiotic EN group (large intestine: 12.516 +/- 1.542) significantly increased as compared with PN group (intestine: 11.189 +/- 2.108; cecum: 10.160 +/- 1.643; P<0.01). The intestinal epithelial tight junctions and microvilli of the probiotic group were more intact than those in the PN group. (3) The bacterial translocations in blood, liver, lung and mesenteric lymph nodes, and the levels of endotoxin were significantly reduced in probiotic (0.082 +/- 0.029) and EN (0.125 +/- 0.040) groups as compared with PN group (0.403 +/- 0.181, P < 0.05).

CONCLUSION

Application of EN combined with probiotics could improve the expression of transmembrane binding proteins (occludin) and IgA, correct the intestinal flora disturbance, maintain gut barrier functions and tight junctions, and reduce the occurrence of gut bacterial translocation.

摘要

目的

探讨肠内营养、肠外营养及经肠道给予益生菌对腹部感染大鼠肠道微生态、上皮紧密连接、免疫及屏障功能的影响。

方法

采用盲肠结扎穿孔法建立大鼠腹部感染模型,将其分为三组:肠外营养组(PN组,n = 7)、肠外营养+肠内营养组(EN组,n = 7)和肠外营养+肠内营养+益生菌组(益生菌组,n = 7),通过空肠造瘘管和颈静脉给予营养支持5天。三组的总营养补充量为等氮和等热量。经空肠造瘘每天给予益生菌10 mL(1×10⁸ cfu/mL)。第6天处死大鼠。培养盲肠粪便中的厌氧菌生长,并用随机扩增多态性DNA分析菌群DNA指纹图谱。采用免疫组织化学方法检测结肠和回肠末端肠上皮浆细胞中的跨膜结合蛋白(闭合蛋白)和IgA水平。通过电子显微镜观察结肠和小肠肠上皮紧密连接的超微结构。培养腔静脉血及肝、肺和肠系膜淋巴结的匀浆组织以测定细菌移位,并检测门静脉血中的内毒素。

结果

(1)EN组和益生菌组的肠道菌群数量高于PN组。EN组和益生菌组的DNA图谱与正常大鼠相似。益生菌组的DNA图谱数量远多于PN组和EN组。此外,PN组有奇怪的条带。(2)与PN组(1.207±0.587,P<0.05,11.189±2.108,P<0.01)相比,EN组(2.309±0.336,15.440±2.383)和益生菌组(2.938±0.515,16.230±3.183)结肠和小肠中闭合蛋白和IgA的表达有所改善。益生菌组中闭合蛋白的表达(肠:2.93±0.515;盲肠:3.40±0.617)高于EN组(肠:2.309±0.336;盲肠:2.076±0.670;P<0.05)。与PN组(肠:11.189±2.108;盲肠:10.160±1.643;P<0.01)相比,EN组(肠:15.440±2.383)和益生菌组(大肠:12.516±1.542)中IgA的表达显著增加。益生菌组的肠上皮紧密连接和微绒毛比PN组更完整。(3)与PN组(0.403±0.181,P<0.05)相比,益生菌组(0.082±0.029)和EN组(0.125±0.040)血、肝、肺和肠系膜淋巴结中的细菌移位及内毒素水平显著降低。

结论

应用EN联合益生菌可改善跨膜结合蛋白(闭合蛋白)和IgA的表达,纠正肠道菌群紊乱,维持肠道屏障功能和紧密连接,减少肠道细菌移位发生。

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