Background

Inflammatory bowel disease occurs when luminal antigens from microbiota stimulate the immune system of an intestinal barrier through defects in the tight junctions lead to an exacerbated response in the genetically predisposed individual. It is well known that the consumption of probiotics improves intestinal health. Specifically, the Lactobacillus sp. strains play a key role in the epithelial intestinal barrier, promote mucin secretion, better performance of tight junctions and prevent apoptosis of epithelial cells. This study aimed to evaluate the role of Kefir (Lactobacillus kefiranofaciens) on inflammation of dextran sodium sulphate (DSS)-induced colitis.

Methods

Forty male Wistar rats were randomised into five groups: Sham; 5% DSS-induced colitis; control Kefir; 5% DSS-induced colitis treated with Kefir; 5% DSS-induced colitis treated with Kefir + 5-ASA (125mg/kg/day) (Ethics Committee on the Use of Animals number 3627290119). The lactic culture of Kefir (Danisco Biolacta, Olstyn, Poland) was administered ad libitum per 6 days. The preparation of the matrix was carried out as follows: whole milk powder was reconstituted to 10% with distilled water and was incubated at 85°C for 15 min using a mechanical shaker. Subsequently, 20 mg freeze-dried Kefir culture was added to 100 ml of the treated milk and fermentation were carried out at 23°C for 16 h. When the desired pH of 4.6 was reached, the fermentation was stopped by cooling the flasks in ice bath and storing them under refrigeration at 4°C until utilisation. The rats were euthanised on day 9. Colonic samples were processed for transmission electron microscope (TEM) and conventional microscopy. Samples were analysed in a TEM EX II 1200 (JEOL, Japan) coupled to a digital camera system GATAN Orius (USA). Histopathological evaluation was performed by a pathologist under a light microscope. Parameters such as inflammation extent, regeneration and crypt damage were graded according to Dieleman et al.³ Statistical analysis was performed by one-way analysis of variance (ANOVA), followed by Tukey’s test using Graph Pad Prism (version 6.0).

Results

Kefir treatment reduces the severity of DSS-induced colitis evidenced by decreased abdominal discomfort and rectal bleeding. Histologically, there was a significant decreased of colonic damage and preservation of goblet cells (p < 0.0001) compared with DSS-colitis control group (Figure 1). The effect of Kefir was also verified by TEM analysis at the ultrastructural level, demonstrating the tight junctions preserved, decreased of reticulum edema and increased of autophagosomes (Figure 2).

Conclusion

Kefir promotes autophagy and protects the intestinal barrier against damage caused by DSS.

中文翻译：

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P056开菲尔（Lactobacillus kefiranofaciens）促进自噬并保护肠屏障免受DSS诱发的结肠炎

背景

当来自微生物群的腔内抗原通过紧密连接的缺陷刺激肠道屏障的免疫系统导致遗传易感个体的反应加剧时，就会发生炎症性肠病。众所周知，食用益生菌可以改善肠道健康。具体而言，乳杆菌属。菌株在上皮小肠屏障中起关键作用，促进粘蛋白分泌，紧密连接的更好性能并防止上皮细胞凋亡。这项研究旨在评估开菲尔（乳酸杆菌kefiranofaciens）在右旋糖酐硫酸钠（DSS）引起的结肠炎炎症中的作用。

方法

将四十只雄性Wistar大鼠随机分为五组：假手术组；假手术组。5％DSS诱发的结肠炎；控制开菲尔; 用开菲尔治疗5％DSS引起的结肠炎；用开菲尔+ 5-ASA（125mg / kg /天）治疗的5％DSS诱导的结肠炎（动物使用伦理委员会编号3627290119）。每6天随意施用开菲尔（Kefir）（Danisco Biolacta，Olstyn，波兰）的乳酸培养物。基质的制备如下：用蒸馏水将全脂奶粉重构至10％，并使用机械振荡器在85°C下孵育15分钟。随后，将20 mg冻干开菲尔培养物添加到100 ml处理过的牛奶中，并在23°C发酵16小时。当达到所需的4.6的pH值时，通过将烧瓶在冰浴中冷却并将其在4°C下冷藏保存直至使用，从而停止发酵。在第9天对大鼠实施安乐死。将结肠样品进行透射电子显微镜（TEM）和常规显微镜检查。在与数码相机系统GATAN Orius（美国）连接的TEM EX II 1200（日本，JEOL）上分析样品。病理学家在光学显微镜下进行组织病理学评估。诸如炎症程度，再生和隐窝损害等参数根据Dieleman等人的方法分级。病理学家在光学显微镜下进行组织病理学评估。诸如炎症程度，再生和隐窝损害等参数根据Dieleman等人的方法分级。病理学家在光学显微镜下进行组织病理学评估。诸如炎症程度，再生和隐窝损害等参数根据Dieleman等人的方法分级。³统计分析通过单向方差分析（ANOVA）进行，然后使用Graph Pad Prism（6.0版）进行Tukey检验。

结果

开菲尔疗法可减轻腹部不适和直肠出血，从而减轻DSS诱发的结肠炎的严重程度。在组织学上，与DSS结肠炎对照组相比，结肠损伤和杯状细胞的保存显着降低（p <0.0001）（图1）。开菲尔的作用还通过超微结构水平的TEM分析得到了证实，表明保留了紧密的连接，网状组织水肿减少，自噬体增加（图2）。

结论

开菲尔可促进自噬并保护肠屏障免受DSS造成的损害。