Hydrogen peroxide activates activator protein-1 and mitogen-activated protein kinases in pancreatic stellate cells
Hydrogen peroxide activates activator protein-1 and mitogen-activated protein kinases in pancreatic stellate cells
复制标题
DOI：
10.1007/s11010-006-9189-4
复制
发表时间：
2006-10-01
影响因子：
4.3
通讯作者：
Shimosegawa, Tooru
中科院分区：
文献类型：
--
作者：
Kikuta, Kazuhiro;Masamune, Atsushi;Shimosegawa, Tooru
激活的胰腺星状细胞（PSC）与胰腺炎症和纤维化的发病机制有关，其中氧化应激被认为发挥着关键作用。活性氧，例如过氧化氢 (H2O2)，可以作为第二信使介导生长因子和细胞因子的作用。但活性氧在 PSC 细胞功能激活和调节中的作用仍然很大程度上未知。我们在此研究了 H2O2 对 PSC 中信号转导途径和细胞功能激活的影响。除非另有说明，PSC 是从雄性 Wistar 大鼠的胰腺中分离出来的，并用于其培养激活的肌成纤维细胞样表型。通过电泳迁移率变动测定和荧光素酶测定检查转录因子的激活。使用抗磷酸特异性抗体通过蛋白质印迹法评估丝裂原激活蛋白（MAP）激酶的激活。评估了 H2O2 对增殖、α(1)(I) 前胶原基因表达和单核细胞趋化蛋白 1 产生的影响。还评估了 H2O2 对培养物中新鲜分离的 PSC 转化的影响。非细胞毒性浓度（高达 100 μM）的 H2O2 会诱导 PSC 中的氧化应激。 H2O2 激活激活蛋白-1，但不激活核因子 kappa B。此外，H2O2 激活三类 MAP 激酶：细胞外信号调节激酶、c-Jun N 末端激酶和 p38 MAP 激酶。 H2O2 诱导 α(1)(I) 前胶原基因表达，但不诱导增殖或单核细胞趋化蛋白 1 的产生。 H2O2 不会引发新鲜分离的 PSC 向肌成纤维细胞样表型的转化。 H2O2 特异性激活这些信号转导途径和胶原蛋白基因表达可能在胰腺纤维化的发病机制中发挥作用。
Activated pancreatic stellate cells (PSCs) are implicated in the pathogenesis of pancreatic inflammation and fibrosis, where oxidative stress is thought to play a key role. Reactive oxygen species such as hydrogen peroxide (H2O2) may act as a second messenger to mediate the actions of growth factors and cytokines. But the role of reactive oxygen species in the activation and regulation of cell functions in PSCs remains largely unknown. We here examined the effects of H2O2 on the activation of signal transduction pathways and cell functions in PSCs. PSCs were isolated from the pancreas of male Wistar rats, and used in their culture-activated, myofibroblast-like phenotype unless otherwise stated. Activation of transcription factors was examined by electrophoretic mobility shift assay and luciferase assay. Activation of mitogen-activated protein (MAP) kinases was assessed by Western blotting using anti-phosphospecific antibodies. The effects of H2O2 on proliferation, alpha(1)(I)procollagen gene expression, and monocyte chemoattractant protein-1 production were evaluated. The effect of H2O2 on the transformation of freshly isolated PSCs in culture was also assessed. H2O2 at non-cytotoxic concentrations (up to 100 mu M) induced oxidative stress in PSCs. H2O2 activated activator protein-1, but not nuclear factor kappa B. In addition, H2O2 activated three classes of MAP kinases: extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 MAP kinase. H2O2 induced alpha(1)(I)procollagen gene expression but did not induce proliferation or monocyte chemoattractant protein-1 production. H2O2 did not initiate the transformation of freshly isolated PSCs to myofibroblast-like phenotype. Specific activation of these signal transduction pathways and collagen gene expression by H2O2 may play a role in the pathogenesis of pancreatic fibrosis.