MAdCAM-1 expression and regulation in murine colonic endothelial cells in vitro
MAdCAM-1 expression and regulation in murine colonic endothelial cells in vitro
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DOI：
10.1097/01.mib.0000160807.53858.1c
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发表时间：
2005-03-01
影响因子：
4.9
通讯作者：
Alexander, JS
中科院分区：
文献类型：
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作者：
Ando, T;Jordan, P;Alexander, JS
背景：虽然粘膜地址素细胞粘附分子-1 (MAdCAM-1) 与炎症性肠病的病因有关，但很少有研究利用结肠前的微血管内皮直接检查 MAdCAM-1。本研究测量了 MAdCAM-1 在新型结肠内皮细胞系 MJC-1 中的表达，以及 MAdCAM-1 的体外调节和功能。 方法：我们使用 ImmortoMice 小鼠（其细胞表达温度敏感的 SV40 大 T 抗原，H-2Kb-tsA58 小鼠）从原代结肠培养物中克隆微血管内皮细胞。通过蛋白质印迹法测定用细胞因子[肿瘤坏死因子(TNF)-α、白细胞介素(IL)-Iβ或干扰素(IFN)-γ]刺激后MAdCAM-1的表达。在细胞因子之前使用药理学阻断剂检查调节 MAdCAM-1 表达的信号路径。我们还使用组成型表达 α4β7 整合素的淋巴细胞检查了淋巴细胞粘附。结果：TNF-α 在 24 小时内以剂量依赖性方式诱导 MAdCAM-1。 MAdCAM-1 诱导依赖于蛋白激酶 C、酪氨酸激酶、p38 丝裂原激活蛋白激酶和核因子 kappa-B/聚腺苷二磷酸核糖聚合酶。 TNF-a 刺激后淋巴细胞粘附增加 2.6 倍，并且在治疗前被抗 MAdCAM-1 抗体抑制（对照与 TNF-a 的 P < 0.05）。结论：在体外，MAdCAM-1 可以通过 TNF-a 刺激诱导油性结肠内皮细胞，可能是研究大肠微血管损伤的有用模型。
Background: Although the mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is associated with the etiology of inflammatory bowel diseases, few studies have directly examined MAdCAM-1 using microvascular endothelium derived front the colon. This study measured the expression of MAdCAM-1 in a novel colon endothelial line MJC-1, as well as MAdCAM-1 regulation and function in vitro.Methods: We cloned microvascular endothelial cells from primary colon cultures using ImmortoMice mice (whose cells express a temperature-sensitive SV40 large T antigen, H-2Kb-tsA58 mice). Expression of MAdCAM-1 after stimulation with cytokines [tumor necrosis factor (TNF)-alpha, interleukin (IL)-Ibeta, or interferon (IFN)-gamma] was determined by Western blotting. Signal paths regulating MAdCAM-1 expression were examined using pharmacological blockers before cytokines. We also examined lymphocyte adhesion using lymphocytes that constitutively express alpha4beta7 integrin.Results: TNF-alpha induced MAdCAM-1 in a dose-dependent manner by 24 hours. MAdCAM-1 induction was protein kinase C, tyrosille kinase, p38 mitogen activated protein kinase, and nuclear-factor kappa-B/poly adenosine diphosphate ribose polymerase dependent. Lymphocyte adhesion was increased 2.6-fold after TNF-a stimulation and was inhibited by anti-MAdCAM-1 antibody before treatment (P < 0.05 control versus TNF-a).Conclusions: in vitro, MAdCAM-1 can be induced oil colon endothelial cells by TNF-a stimulation and may represent a useful model to study microvascular injury in the large intestine.