Rho kinase blockade prevents inflammation via nuclear factor κB inhibition:: Evidence in Crohn's disease and experimental colitis
Rho kinase blockade prevents inflammation via nuclear factor κB inhibition:: Evidence in Crohn's disease and experimental colitis
复制标题
DOI：
10.1016/s0016-5085(03)00283-x
复制
发表时间：
2003-05-01
期刊：
影响因子：
29.4
通讯作者：
Loirand, G
中科院分区：
文献类型：
--
作者：
Segain, JP;de la Blétière, DR;Loirand, G
背景与目的：Rho蛋白参与多种细胞功能的调控。体外研究数据表明RhoA可能参与炎症反应。我们研究了RhoA及其下游效应物Rho激酶在肠道炎症中的作用。方法：采用下拉法测定RhoA的活化情况。利用Rho激酶的特异性抑制剂Y-27632，通过分子生物学、免疫学和生化方法研究了Rho激酶在体内和体外炎症反应中的作用。结果：克罗恩病患者和2,4,6-三硝基苯磺酸致结肠炎大鼠炎症性肠黏膜RhoA活化增加。大鼠口服Y-27632可明显减轻结肠炎症。在体外，仅激活RhoA就足以诱导肿瘤坏死因子的产生。Y-27632抑制固有层和外周血单核细胞产生肿瘤坏死因子- α和白细胞介素-1 β。Rho激酶抑制阻止核因子kappaB激活和I-kappaB磷酸化和降解。我们发现Rho激酶与I-kappaB激酶α结合并激活，Y-27632阻止I-kappaB激酶的激活。结论：我们的研究首次提供了Rho激酶激活I-kappaB激酶，从而激活核因子kappaB的证据，表明Rho激酶在炎症反应和肠道炎症中起关键作用。特异性抑制Rho激酶可能是治疗克罗恩病患者的一种有希望的方法。
Background & Aims: Rho proteins are involved in the regulation of several cellular functions. Data from in vitro studies suggest that RhoA could be involved in the inflammatory response. We investigated the role of RhoA and its downstream effector Rho kinase in intestinal inflammation. Methods: Activation of RhoA was assessed by pull-down assays. A specific inhibitor of Rho kinase, Y-27632, was used to examine the role of Rho kinase in inflammatory response in vivo and in vitro by molecular biology and by immunological and biochemical approaches. Results: Increased activation of RhoA was found in inflamed intestinal mucosa of patients with Crohn's disease and of rats with 2,4,6-trinitrobenzene sulfonic acid-induced colitis. Oral administration of Y-27632 in rats significantly reduced the colonic inflammation. In vitro, activation of RhoA alone was sufficient to induce tumor necrosis factor production. Y-27632 inhibited production of tumor necrosis factor-alpha and interleukin-1beta by lamina propria and peripheral blood mononuclear cells. Rho kinase inhibition prevented nuclear factor kappaB activation and I-kappaB phosphorylation and degradation. We showed that Rho kinase associates with and activates I-kappaB kinase alpha and that Y-27632 prevents I-kappaB kinase activation. Conclusions: Our study provides the first evidence that Rho kinase activates I-kappaB kinase and, thus, nuclear factor kappaB, suggesting a key role of Rho kinase in inflammatory responses and intestinal inflammation. Specific inhibition of Rho kinase may be a promising approach for the treatment of patients with Crohn's disease.