Butyrate Enhances the Intestinal Barrier by Facilitating Tight Junction Assembly via Activation of AMP-Activated Protein Kinase in Caco-2 Cell Monolayers
Butyrate Enhances the Intestinal Barrier by Facilitating Tight Junction Assembly via Activation of AMP-Activated Protein Kinase in Caco-2 Cell Monolayers
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DOI：
10.3945/jn.109.104638
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发表时间：
2009-09-01
影响因子：
4.2
通讯作者：
Lin, Jing
中科院分区：
文献类型：
--
作者：
Peng, Luying;Li, Zhong-Rong;Lin, Jing
丁酸盐是SCFA中的一种，可促进肠道屏障的发育。然而，丁酸盐调节肠屏障的分子机制尚不清楚。为了检验丁酸盐对肠屏障的作用是通过调节涉及AMP活化蛋白激酶（AMPK）活化的紧密连接组装来介导的这一假设，我们通过测量Caco-2细胞单层模型中的跨上皮电阻（TER）和菊粉渗透性来确定丁酸盐对肠屏障的作用。我们进一步使用钙开关分析来研究上皮紧密连接的组装，并确定丁酸盐对上皮紧密连接的组装和AMPK活性的影响。我们证明丁酸盐处理增加AMPK活性，并加速紧密连接的组装，如紧密连接蛋白的重组以及TER的发展所示。在钙开关诱导的紧密连接组装过程中，丁酸盐也上调AMPK活性。化合物C，一种特异性AMPK抑制剂，抑制丁酸诱导的AMPK活化。丁酸盐对标准培养基中以及钙转换后TER增加的促进作用被化合物C消除。我们得出结论，丁酸盐通过调节紧密连接的组装来增强肠屏障。这一动态过程由AMPK的激活介导。这些结果表明，SCFA和细胞内能量传感器之间的一个有趣的联系，为肠道屏障的发展。J.营养139：1619-1625，2009.
Butyrate, one of the SCFA, promotes the development of the intestinal barrier. However, the molecular mechanisms underlying the butyrate regulation of the intestinal barrier are unknown. To test the hypothesis that the effect of butyrate on the intestinal barrier is mediated by the regulation of the assembly of tight junctions involving the activation of the AMP-activated protein kinase (AMPK), we determined the effect of butyrate on the intestinal barrier by measuring the transepithelial electrical resistance (TER) and inulin permeability in a Caco-2 cell monolayer model. We further used a calcium switch assay to study the assembly of epithelial tight junctions and determined the effect of butyrate on the assembly of epithelial tight junctions and AMPK activity, We demonstrated that the butyrate treatment increased AMPK activity and accelerated the assembly of tight junctions as shown by the reorganization of tight junction proteins, as well as the development of TER. AMPK activity was also upregulated by butyrate during calcium switch-induced tight junction assembly. Compound C, a specific AMPK inhibitor, inhibited the butyrate-induced activation of AMPK. The facilitating effect of butyrate on the increases in TER in standard culture media, as well as after calcium switch, was abolished by compound C. We conclude that butyrate enhances the intestinal barrier by regulating the assembly of tight junctions. This dynamic process is mediated by the activation of AMPK. These results suggest an intriguing link between SCFA and the intracellular energy sensor for the development of the intestinal barrier. J. Nutr. 139: 1619-1625, 2009.