Protective Effect of Kaempferol on LPS-Induced Inflammation and Barrier Dysfunction in a Coculture Model of Intestinal Epithelial Cells and Intestinal Microvascular Endothelial Cells
Protective Effect of Kaempferol on LPS-Induced Inflammation and Barrier Dysfunction in a Coculture Model of Intestinal Epithelial Cells and Intestinal Microvascular Endothelial Cells
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山奈酚对肠上皮细胞与肠微血管内皮细胞共培养模型中LPS诱导的炎症和屏障功能障碍的保护作用
DOI：
10.1021/acs.jafc.9b06294
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发表时间：
2020-01-08
影响因子：
6.1
通讯作者：
Zhang, Bingkun
中科院分区：
文献类型：
--
作者：
Bian, Yifei;Dong, Yuanyang;Zhang, Bingkun
炎症性肠病（Inflammatory bowel disease，IBD）是一种肠粘膜和粘膜下层的慢性炎症性疾病，其特征是肠上皮屏障破坏、炎症介质产生增加和过度组织损伤。肠上皮细胞以及微血管内皮细胞在IBD中起重要作用。为了研究山萘酚在IBD发病过程中的作用，我们建立了一种新的上皮内皮细胞共培养模型，以研究肠道炎症和屏障功能。数据表明，跨上皮电阻（TEER）明显增加，（1222 +/- 60.40 Omega cm 2 vs 1371 +/- 38.77 Omega cm 2），FITC通量降低（180.8 +/- 20.06 μ g/mL vs 136.7 +/- 14.78 μ g/mL），并在与内皮细胞特异性共培养的Caco-2中上调occludin和claudin-2表达。同时，80 μ M山奈酚可减轻1 μ g/mL脂多糖（LPS）诱导的TEER下降、FITC通量增加和白细胞介素-8（IL-8）过表达。此外，山奈酚还改善了LPS诱导的闭合小带-1（Z 0 -1）、闭合蛋白和封闭蛋白-2蛋白表达的降低，以及LPS诱导的NF-κ B和I-κ B磷酸化水平的蛋白表达的抑制。我们的研究结果表明，山奈酚通过抑制NF-κ B B信号通路的激活，减轻了LPS诱导的上皮-内皮共培养模型中Caco-2单层细胞的IL-8分泌和屏障功能障碍。
Inflammatory bowel disease (IBD) is a chronic inflammatory disease of intestinal mucosa and submucosa, characterized by the disruption of the intestinal epithelial barrier, increased production of inflammatory mediators, and excessive tissue injury. Intestinal epithelial cells, as well as microvascular endothelial cells, play important roles in IBD. To study the potential effects of kaempferol in IBD progress, we established a novel epithelial endothelial cells coculture model to investigate the intestinal inflammation and barrier function. Data demonstrated an obvious increased transepithelial electrical resistance (TEER) (1222 +/- 60.40 Omega cm2 vs 1371 +/- 38.77 Omega cm(2)), decreased flux of FITC (180.8 +/- 20.06 mu g/mL vs 136.7 +/- 14.78 mu g/mL), and up-regulated occludin and claudin-2 expression in Caco-2 that was specifically cocultured with endothelial cells. Meanwhile, 80 mu M kaempferol alleviated the drop of TEER, the increase of FITC flux, and the overexpression of interleukin-8 (IL-8) induced by 1 mu g/mL lipopolysaccharide (LPS). Additionally, kaempferol also ameliorated the LPS-induced decrease of protein expression of zonula occludens-1 (Z0-1), occludin, and claudin-2, together with the inhibited protein expressions of the phosphorylation level of NF-kappa B and I-kappa B induced by LPS. Our results suggest that kaempferol alleviates the IL-8 secretion and barrier dysfunction of the Caco-2 monolayer in the LPS-induced epithelial-endothelial coculture model via inhibiting the NF-kappa B signaling pathway activation.