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罗伊氏乳杆菌I5007在脂多糖刺激下对IPEC-J2细胞以及在正常条件下对新生仔猪的紧密连接蛋白表达具有调节作用。

Lactobacillus reuteri I5007 modulates tight junction protein expression in IPEC-J2 cells with LPS stimulation and in newborn piglets under normal conditions.

作者信息

Yang Fengjuan, Wang Aina, Zeng Xiangfang, Hou Chengli, Liu Hong, Qiao Shiyan

机构信息

State Key Laboratory of Animal Nutrition, China Agricultural University, Beijing, 100193, China.

Weifang Business Vocational College, Zhucheng, 262234, China.

出版信息

BMC Microbiol. 2015 Feb 15;15:32. doi: 10.1186/s12866-015-0372-1.

Abstract

BACKGROUND

Tight junctions (TJs) maintain the intestinal mucosal barrier, dysfunction of which plays a vital role in the pathophysiology of a variety of gastrointestinal disorders. Previously, we have shown that L. reuteri I5007 maintained the gut epithelial barrier in newborn piglets. Here we aimed to decipher the influence of L. reuteri I5007 on tight junction (TJ) protein expression both in vivo and in vitro.

RESULTS

We found that L. reuteri I5007 significantly increased the protein abundance of intestinal epithelial claudin-1, occludin and zonula occluden-1 (ZO-1) in newborn piglets (orally administrated with 6 × 10(9) CFU of L. reuteri I5007 daily for 14 days). In vitro, treatment with L. reuteri I5007 alone maintained the transepithelial electrical resistance (TEER) of IPEC-J2 cells with time. In addition, IPEC-J2 cells were stimulated with 1 μg/mL lipopolysaccharide (LPS) for 1, 4, 8, 12 or 24 h, following pre-treatment with L. reuteri I5007 or its culture supernatant for 2 h. The results showed that LPS time-dependently induced (significantly after 4 or 8 h) the expression of TNF-α and IL-6, and decreased TJ proteins, which was reversed by pre-treatment of L. reuteri I5007 or its culture supernatant.

CONCLUSIONS

L. reuteri I5007 had beneficial effects on the expression of TJ proteins in newborn piglets and the in-vitro results showed this strain had a positive effect on TEER of cells and inhibited the reduction of TJ proteins expression induced by LPS. These findings indicated L. reuteri I5007 may have potential roles in protection TJ proteins in TJ-deficient conditions.

摘要

背景

紧密连接(TJs)维持肠道黏膜屏障,其功能障碍在多种胃肠道疾病的病理生理学中起重要作用。此前,我们已表明罗伊氏乳杆菌I5007可维持新生仔猪的肠道上皮屏障。在此,我们旨在阐明罗伊氏乳杆菌I5007在体内和体外对紧密连接(TJ)蛋白表达的影响。

结果

我们发现罗伊氏乳杆菌I5007显著增加了新生仔猪肠道上皮紧密连接蛋白-1、闭合蛋白和闭合小环蛋白-1(ZO-1)的蛋白丰度(每天口服6×10⁹CFU罗伊氏乳杆菌I5007,持续14天)。在体外,单独用罗伊氏乳杆菌I5007处理可随时间维持IPEC-J2细胞的跨上皮电阻(TEER)。此外,在用罗伊氏乳杆菌I5007或其培养上清预处理2小时后,用1μg/mL脂多糖(LPS)刺激IPEC-J2细胞1、4、8、12或24小时。结果表明,LPS可时间依赖性地诱导(4或8小时后显著)肿瘤坏死因子-α和白细胞介素-6的表达,并降低TJ蛋白,而罗伊氏乳杆菌I5007或其培养上清的预处理可逆转这一现象。

结论

罗伊氏乳杆菌I5007对新生仔猪TJ蛋白的表达具有有益作用,体外结果表明该菌株对细胞的TEER有积极影响,并抑制了LPS诱导的TJ蛋白表达降低。这些发现表明罗伊氏乳杆菌I5007在TJ缺陷条件下可能对保护TJ蛋白具有潜在作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcf0/4350629/c85c4b62cb0f/12866_2015_372_Fig1_HTML.jpg

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