Protective effects of carbon monoxide-releasing molecule-2 on the barrier function of intestinal epithelial cells.
Protective effects of carbon monoxide-releasing molecule-2 on the barrier function of intestinal epithelial cells.
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一氧化碳释放分子 2 对肠上皮细胞屏障功能的保护作用。
DOI：
10.1371/journal.pone.0104032
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Wang G
中科院分区：
文献类型：
--
作者：
Mu X;Pan C;Zheng S;Alhamdi Y;Sun B;Shi Q;Wang X;Sun Z;Toh C;Wang G
探讨一氧化碳释放分子-2（CORM-2）对肠上皮细胞屏障功能的保护作用及其机制。用CORM-2预孵育1小时后，用50 μg/ml脂多糖（LPS）刺激培养的肠上皮IEC-6细胞。采用ELISA试剂盒检测培养液中细胞因子水平。用Millipore电阻系统（ERS-2; Millipore）测量Transwell中IEC-6细胞单层的跨上皮电阻（TER），并计算为LPS处理后不同时间点的Ω/cm 2。还使用FITC-葡聚糖测量渗透性变化。用特异性抗体免疫印迹法检测紧密连接蛋白（紧密连接蛋白和ZO-1）和肌球蛋白轻链（MLC）磷酸化水平。随后的结构变化的TJ可视化使用透射电子显微镜（TEM）。CORM-2显著降低LPS诱导的TNF-α和IL-1β的分泌。CORM-2浓度依赖性地抑制LPS诱导的TER降低和对FITC-dextran通透性的增加（P<0.05）。LPS诱导的紧密连接蛋白的减少和MLC磷酸化的增加也减弱。在LPS处理的细胞，TEM显示减少电子致密材料和中断的TJ和桥粒之间的顶侧边缘相邻的细胞，这是防止CORM-2治疗。本研究表明CORM-2作为一种新型的CO释放分子，具有保护LPS刺激的肠上皮细胞屏障功能的能力。抑制炎性细胞因子释放、恢复TJ蛋白和抑制MLC磷酸化是CORM-2的保护作用之一。
To investigate the protective effects and mechanisms of carbon monoxide-releasing molecule-2 (CORM-2) on barrier function of intestinal epithelial cells. After pre-incubation with CORM-2 for 1 hour, cultured intestinal epithelial IEC-6 cells were stimulated with 50 µg/ml lipopolysaccharides (LPS). Cytokines levels in culture medium were detected using ELISA kits. Trans-epithelial electrical resistance (TER) of IEC-6 cell monolayers in Transwells were measured with a Millipore electric resistance system (ERS-2; Millipore) and calculated as Ω/cm2 at different time points after LPS treatment. The permeability changes were also measured using FITC-dextran. The levels of tight junction (TJ) proteins (occludin and ZO-1) and myosin light chain (MLC) phosphorylation were detected using Western blotting with specific antibodies. The subsequent structural changes of TJ were visualized using transmission electron microscopy (TEM). CORM-2 significantly reduced LPS-induced secretion of TNF-α and IL-1β. The LPS-induced decrease of TER and increase of permeability to FITC-dextran were inhibited by CORM-2 in a concentration dependent manner (P<0.05). LPS-induced reduction of tight junction proteins and increase of MLC phosphorylation were also attenuated. In LPS-treated cells, TEM showed diminished electron-dense material and interruption of TJ and desmosomes between the apical lateral margins of adjoining cells, which were prevented by CORM-2 treatment. The present study demonstrates that CORM-2, as a novel CO-releasing molecule, has ability to protect the barrier function of LPS-stimulated intestinal epithelial cells. Inhibition of inflammatory cytokines release, restoration of TJ proteins and suppression of MLC phosphorylation are among the protective effects of CORM-2.
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影响因子：
4.8
作者：
Lee, TS;Tsai, HL;Chau, LY
通讯作者：
Chau, LY
影响因子：
7.3
作者：
Megias, J.;Busserolles, J.;Alcaraz, M. J.
通讯作者：
Alcaraz, M. J.
影响因子：
3.7
作者：
García-Arnandis I;Guillén MI;Gomar F;Castejón MA;Alcaraz MJ
通讯作者：
Alcaraz MJ
影响因子：
5.1
作者：
Katada, Kazuhiro;Bihari, Aurelia;Cepinskas, Gediminas
通讯作者：
Cepinskas, Gediminas
影响因子：
3.7
作者：
Guo, Ying;Ramachandran, Charanya;Srinivas, Sangly P.
通讯作者：
Srinivas, Sangly P.