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布拉氏酵母菌对葡聚糖硫酸钠诱导的结肠炎小鼠肠黏膜屏障的保护作用。

Protective effect of Saccharomyces boulardii on intestinal mucosal barrier of dextran sodium sulfate-induced colitis in mice.

机构信息

Department of Gastroenterology, Peking University First Hospital, Beijing 100034, China.

Department of Urology, Peking University First Hospital, Beijing 100034, China.

出版信息

Chin Med J (Engl). 2019 Aug 20;132(16):1951-1958. doi: 10.1097/CM9.0000000000000364.

Abstract

BACKGROUND

The effect and mechanism of Saccharomyces boulardii (Sb) in inflammatory bowel disease are unclear. The objective of the study was to evaluate the impact of Sb on intestinal mucosal barrier and intestinal flora in a colitis mouse model.

METHODS

Forty C57BL/6J male mice were randomly assigned to five groups: normal control group (A), pathologic control group (B), Sb treatment group (C), mesalazine treatment group (D), and Sb combined with mesalazine treatment group (E). Colitis was induced by the addition of 2.5% (wt/vol) dextran sodium sulfate (DSS) in the drinking water ad libitum for 7 days. The general condition, weight change, stool property, and bloody stool level of mice were observed to evaluate the disease activity index. The expression of zona occludens-1 (ZO-1) and occludin in intestinal tissue were measured by immunohistochemistry. The level of tumor necrosis factor-α (TNF-α) and interleukin (IL)-8 in plasma was measured by enzyme linked immunosorbent assay. Inter-cellular tight junctions were observed by transmission electron microscopy. The feces and intestinal contents were collected sterilely, and intestinal flora was analyzed by 16S rRNA sequencing.

RESULTS

Compared with group B, Sb reduced the disease activity index and histological score of group C (disease activity index: group B 2.708 ± 0.628, group C 1.542 ± 0.616, PBC = 0.005; histological score: group B 9.875 ± 3.271, group C 4.750 ± 1.832, PBC = 0.005) in DSS-induced colitis in mice. Sb exerted a protect effect on the expression of ZO-1 (group B 2.075 ± 1.176, group C 4.225 ± 1.316, PBC = 0.019) and occludin (group B 2.200 ± 0.968, group C 3.525 ± 1.047, PBC = 0.023). Compared with group B, Sb decreased the level of TNF-α and IL-8 of group C (TNF-α: group B 716.323 ± 44.691 ng/L, group C 521.740 ± 90.121 ng/L, PBC = 0.001; IL-8: group B 128.992 ± 11.475 pg/mL, group C 106.283 ± 15.906 pg/mL, PBC = 0.012). Treatment with Sb preserved the tight junctions and ameliorated microvilli and inter-cellular space. Treatment with Sb also showed its own characteristics: a higher percentage of Bacteroidetes and a lower percentage of Firmicutes, with significant differences or a significant trend. The proportion of the S24-7 family was increased significantly in the Sb treatment group.

CONCLUSIONS

Sb shows an anti-inflammatory effect and has a protective effect on the intestinal mucosal mechanical barrier. Sb may up-regulate the abundance of family S24-7 specifically, and maybe a mechanism underlying its function.

摘要

背景

酿酒酵母(Sb)在炎症性肠病中的作用和机制尚不清楚。本研究旨在评估 Sb 对结肠炎小鼠模型肠黏膜屏障和肠道菌群的影响。

方法

将 40 只 C57BL/6J 雄性小鼠随机分为五组:正常对照组(A)、病理对照组(B)、Sb 治疗组(C)、美沙拉嗪治疗组(D)和 Sb 联合美沙拉嗪治疗组(E)。通过在饮用水中添加 2.5%(wt/vol)葡聚糖硫酸钠(DSS),使小鼠自由饮用,持续 7 天诱导结肠炎。观察小鼠的一般情况、体重变化、粪便性状和血便程度,以评估疾病活动指数。采用免疫组织化学法检测肠组织中闭锁蛋白-1(ZO-1)和闭合蛋白的表达。采用酶联免疫吸附试验检测血浆中肿瘤坏死因子-α(TNF-α)和白细胞介素(IL)-8 的水平。采用透射电镜观察细胞间紧密连接。无菌采集粪便和肠内容物,采用 16S rRNA 测序分析肠道菌群。

结果

与 B 组相比,C 组的疾病活动指数和组织学评分均降低(疾病活动指数:B 组 2.708 ± 0.628,C 组 1.542 ± 0.616,PBC=0.005;组织学评分:B 组 9.875 ± 3.271,C 组 4.750 ± 1.832,PBC=0.005)。Sb 对 ZO-1(B 组 2.075 ± 1.176,C 组 4.225 ± 1.316,PBC=0.019)和闭合蛋白(B 组 2.200 ± 0.968,C 组 3.525 ± 1.047,PBC=0.023)的表达有保护作用。与 B 组相比,C 组的 TNF-α和 IL-8 水平降低(TNF-α:B 组 716.323 ± 44.691 ng/L,C 组 521.740 ± 90.121 ng/L,PBC=0.001;IL-8:B 组 128.992 ± 11.475 pg/mL,C 组 106.283 ± 15.906 pg/mL,PBC=0.012)。Sb 治疗组可维持紧密连接,改善微绒毛和细胞间隙。Sb 治疗组还表现出其自身的特点:拟杆菌门的比例较高,厚壁菌门的比例较低,具有显著差异或显著趋势。Sb 治疗组 S24-7 家族的比例显著增加。

结论

Sb 具有抗炎作用,对肠黏膜机械屏障具有保护作用。Sb 可能通过特异性地上调家族 S24-7 的丰度来发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bfff/6708699/e6bee38d4a4a/cm9-132-1951-g001.jpg

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