Probiotic bacteria regulate intestinal epithelial permeability in experimental ileitis by a TNF-dependent mechanism.
Probiotic bacteria regulate intestinal epithelial permeability in experimental ileitis by a TNF-dependent mechanism.
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DOI：
10.1371/journal.pone.0042067
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Pizarro TT
中科院分区：
文献类型：
--
作者：
Corridoni D;Pastorelli L;Mattioli B;Locovei S;Ishikawa D;Arseneau KO;Chieppa M;Cominelli F;Pizarro TT
我们先前表明，益生菌混合物VSL#3可预防SAMP/YitFc（SAMP）小鼠回肠炎发作，该作用与刺激上皮源性TNF相关。本研究旨在确定VSLI #3介导的上皮屏障功能保护机制，并进一步研究TNF在预防SAMP回肠炎中的“矛盾”作用。在SAMP小鼠中，在炎症发作前和确立疾病期间，通过测量暴露于VSLI #3条件培养基（CM）、TNF或VSLI #3-CM +抗TNF后离体培养回肠的跨上皮电阻（TEER），评价渗透性。通过qRT-PCR、蛋白质印迹和共聚焦显微镜评估紧密连接（TJ）蛋白，并在来自SAMP和对照AKR小鼠的新鲜分离的肠上皮细胞（IEC）中测量TNFRI/TNFRII表达。VSLI #3-CM或TNF培养导致炎症前SAMP的回肠细胞旁通透性降低，但对已确诊疾病的SAMP无影响，而添加抗TNF可消除这些作用。在用VSL#3-CM或TNF刺激后，发生TJ蛋白、密蛋白-2和occludin的调节，密蛋白-2显著降低，而occludin增加。与溶剂相比，与VSL#3-CM孵育的SAMP回肠上清液中TNF蛋白水平升高，而IEC衍生的TNFR mRNA表达在幼龄SAMP小鼠中降低，在炎症SAMP小鼠中升高。我们的数据表明，先前确定的VSLI #3预防SAMP回肠炎的疗效是由于TNF对肠上皮的直接先天性和稳态作用、TJ蛋白、claudin-2和occludin的调节以及肠通透性的总体改善。
We previously showed that the probiotic mixture, VSL#3, prevents the onset of ileitis in SAMP/YitFc (SAMP) mice, and this effect was associated with stimulation of epithelial-derived TNF. The aim of this study was to determine the mechanism(s) of VSL#3-mediated protection on epithelial barrier function and to further investigate the “paradoxical” effects of TNF in preventing SAMP ileitis. Permeability was evaluated in SAMP mice prior to the onset of inflammation and during established disease by measuring transepithelial electrical resistance (TEER) on ex vivo-cultured ilea following exposure to VSL#3 conditioned media (CM), TNF or VSL#3-CM + anti-TNF. Tight junction (TJ) proteins were assessed by qRT-PCR, Western blot, and confocal microscopy, and TNFRI/TNFRII expression measured in freshly isolated intestinal epithelial cells (IEC) from SAMP and control AKR mice. Culture with either VSL#3-CM or TNF resulted in decreased ileal paracellular permeability in pre-inflamed SAMP, but not SAMP with established disease, while addition of anti-TNF abrogated these effects. Modulation of the TJ proteins, claudin-2 and occludin, occurred with a significant decrease in claudin-2 and increase in occludin following stimulation with VSL#3-CM or TNF. TNF protein levels increased in supernatants of SAMP ilea incubated with VSL#3-CM compared to vehicle, while IEC-derived TNFR mRNA expression decreased in young, and was elevated in inflamed, SAMP versus AKR mice. Our data demonstrate that the previously established efficacy of VSL#3 in preventing SAMP ileitis is due to direct innate and homeostatic effects of TNF on the gut epithelium, modulation of the TJ proteins, claudin-2 and occludin, and overall improvement of intestinal permeability.
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DOI：
10.1152/ajpregu.00470.2005
发表时间：
2006-06-01
影响因子：
2.8
作者：
Günzel, D;Florian, P;Gitter, AH
通讯作者：
Gitter, AH
通过表达突变型紧密连接膜蛋白，细胞旁通透性和跨上皮电阻的功能性分离以及顶端-基底外侧膜内扩散屏障的破坏。
影响因子：
7.8
作者：
Balda, M S;Whitney, J A;Flores, C;Gonzalez, S;Cereijido, M;Matter, K
通讯作者：
Matter, K
影响因子：
3.1
作者：
Ebach, DR;Riehl, TE;Stenson, WF
通讯作者：
Stenson, WF
DOI：
10.1152/ajpgi.90227.2008
发表时间：
2008-11-01
影响因子：
4.5
作者：
Ewaschuk, Julia B.;Diaz, Hugo;Madsen, Karen L.
通讯作者：
Madsen, Karen L.
影响因子：
6.8
作者：
Chapman, Therese M.;Plosker, Greg L.;Figgitt, David P.
通讯作者：
Figgitt, David P.