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在葡聚糖硫酸钠诱导的小鼠结肠炎模型中,丁酸梭菌通过维持紧密连接蛋白表达和调节微生物群来保护上皮屏障。

Clostridium butyricum protects the epithelial barrier by maintaining tight junction protein expression and regulating microflora in a murine model of dextran sodium sulfate-induced colitis.

作者信息

Li Hui, Gong Yi, Xie Ying, Sun Qian, Li Yan

机构信息

a Department of Gastroenterology , Shengjing Hospital Affiliated to China Medical University , Sanhao Street 36 , Shenyang , Liaoning , PR China.

出版信息

Scand J Gastroenterol. 2018 Sep;53(9):1031-1042. doi: 10.1080/00365521.2016.1192678. Epub 2018 Aug 24.

Abstract

OBJECTIVE

To investigate the protective effects of Clostridium butyricum on the epithelial barrier in dextran sodium sulfate (DSS)-induced colitis.

METHODS

Eight-week-old BALB/c mice were divided into a healthy control group and DSS-induced groups, including negative control (normal saline), 5-aminosalicylic acid (5-ASA), C. butyricum group, and 5-ASA + C. butyricum groups. Colitis was induced by 5% DSS ad libitum for 7 d. We assessed the disease activity index (DAI), histological grading, and ultrastructural changes by transmission electron microscopy. Stools were collected for bacterial microflora analysis. Tight junction (TJ) proteins, including claudin-1, claudin-2, occluding, and zonula occludens-1, were detected by immunohistochemical staining and western blot. We also assessed NF-κB and cytokines, including IL-1β, IL-13, and IL-10, by western blot.

RESULTS

C. butyricum decreased DAI and histological injury scores in DSS-induced mice, and repaired the damaged structure of TJs. Moreover, C. butyricum exerted its regulatory effect on fecal microflora by increasing and decreasing the growth of Lactobacillus spp. and Enterococcus spp., respectively, in the colon lumen. Expression levels of claudin-1, occludin and zonula occludens-1 were also elevated by the administration of C. butyricum. In addition, C. butyricum increased the expression of the anti-inflammatory cytokine, IL-10, and decreased levels of pro-inflammatory cytokines, including IL-1β, TNF, and IL-13, as well as NF-κB. Moreover, the beneficial effects of C. butyricum combined with 5-ASA were superior to treatment with C. butyricum only.

CONCLUSION

In a mouse model of ulcerative colitis (UC), C. butyricum exerted a protective effect on the epithelial barrier by regulating microflora, maintaining the expression of TJ proteins and exerting immunoregulatory effects.

摘要

目的

探讨丁酸梭菌对葡聚糖硫酸钠(DSS)诱导的结肠炎上皮屏障的保护作用。

方法

将8周龄的BALB/c小鼠分为健康对照组和DSS诱导组,包括阴性对照组(生理盐水)、5-氨基水杨酸(5-ASA)组、丁酸梭菌组和5-ASA +丁酸梭菌组。通过自由饮用5% DSS诱导结肠炎7天。我们通过透射电子显微镜评估疾病活动指数(DAI)、组织学分级和超微结构变化。收集粪便进行细菌微生物区系分析。通过免疫组织化学染色和蛋白质印迹法检测紧密连接(TJ)蛋白,包括闭合蛋白-1、闭合蛋白-2、封闭蛋白和紧密连接蛋白-1。我们还通过蛋白质印迹法评估核因子κB(NF-κB)和细胞因子,包括白细胞介素-1β(IL-1β)、白细胞介素-13(IL-13)和白细胞介素-10(IL-10)。

结果

丁酸梭菌降低了DSS诱导小鼠的DAI和组织学损伤评分,并修复了受损的TJ结构。此外,丁酸梭菌通过分别增加和减少结肠腔内乳酸杆菌属和肠球菌属的生长,对粪便微生物区系发挥调节作用。给予丁酸梭菌还可提高闭合蛋白-1、封闭蛋白和紧密连接蛋白-1的表达水平。此外,丁酸梭菌增加了抗炎细胞因子IL-10的表达,降低了促炎细胞因子(包括IL-1β、肿瘤坏死因子(TNF)和IL-13)以及NF-κB的水平。此外,丁酸梭菌与5-ASA联合使用的有益效果优于单独使用丁酸梭菌治疗。

结论

在溃疡性结肠炎(UC)小鼠模型中,丁酸梭菌通过调节微生物区系、维持TJ蛋白的表达和发挥免疫调节作用,对上皮屏障发挥保护作用。

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