Inhibition of p38 Mitogen-Activated Protein Kinase Attenuates Butyrate-Induced Intestinal Barrier Impairment in a Caco-2 Cell Monolayer Model
Inhibition of p38 Mitogen-Activated Protein Kinase Attenuates Butyrate-Induced Intestinal Barrier Impairment in a Caco-2 Cell Monolayer Model
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DOI：
10.1097/mpg.0000000000000369
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发表时间：
2014-08-01
影响因子：
2.9
通讯作者：
Lin, Jing
中科院分区：
文献类型：
--
作者：
Huang, Xiao-Zhong;Li, Zhong-Rong;Lin, Jing
目的：丁酸盐可诱导分化中的肠上皮细胞凋亡。本研究旨在探讨p38丝裂原活化蛋白激酶（MAPK）在丁酸诱导的肠屏障损伤中的作用。方法：采用Caco-2细胞单层模型，通过测量跨上皮电阻（TER）来确定肠屏障。通过测量荧光素异硫氰酸酯缀合的菊粉（菊粉-FITC）的跨上皮通道来确定渗透性。用扫描电子显微镜检查单分子膜的形态。Annexin V FITC标记和流式细胞术检测细胞凋亡状态。结果：5 mM丁酸可增加Caco-2细胞的凋亡率，并通过降低TER和增加菊糖-FITC的通透性来诱导肠屏障功能的损害。丁酸盐处理以浓度和时间依赖性方式激活p38 MAPK。SB 203580是一种特异性p38抑制剂，可抑制丁酸盐诱导的Caco-2细胞凋亡。SB 203580可明显减轻丁酸盐诱导的Caco-2细胞单层屏障功能损害。结论：丁酸盐可激活p38 MAPK，并参与丁酸盐诱导的细胞凋亡和肠屏障功能损害。抑制p38 MAPK可显著减轻丁酸诱导的肠屏障功能障碍。
Objectives: Butyrate is well known to induce apoptosis in differentiating intestinal epithelial cells. The present study was designed to examine the role of p38 mitogen-activated protein kinase (MAPK) in butyrate-induced intestinal barrier impairment.Methods: The intestinal barrier was determined by measuring the transepithelial electrical resistance (TER) in a Caco-2 cell monolayer model. The permeability was determined by measuring transepithelial passage of fluorescein isothiocyanate-conjugated inulin (inulin-FITC). The morphology of the monolayers was examined with scanning electron microscopy. The apoptosis status was determined by annexin V FITC labeling and flow cytometry. The activity of p38 MAPK was determined by the phosphorylation status of p38 with Western blotting.Results: Butyrate at 5 mM increases the apoptosis rate of Caco-2 cells and induces impairment of intestinal bather functions as determined by decreased TER and increased inulin-FITC permeability. Butyrate treatment activates p38 MAPK in a concentration- and time-dependent manner. SB203580, a specific p38 inhibitor, inhibits butyrate-induced Caco-2 cell apoptosis. Treatment of SB203580 significantly attenuates the butyrate-induced impairment of barrier functions in the Caco-2 cell monolayer model.Conclusions: p38 MAPK can be activated by butyrate and is involved in the butyrate-induced apoptosis and impairment of intestinal barrier function. Inhibition of p38 MAPK can significantly attenuate butyrate-induced intestinal barrier dysfunction.