Surgical Services, Iowa City Veterans Affairs Medical Center, and Department of Surgery, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, Iowa, USA.
Pancreatology. 2010;10(2-3):119-28. doi: 10.1159/000290656. Epub 2010 May 7.
The role of the p38 mitogen-activated protein (MAP) kinase in acute pancreatitis pathogenesis is controversial. We hypothesize that p38 plays a role in regulating NF-kappaB activation in exocrine pancreatic cells.
AR42J cells incorporating an NF-kappaB-responsive luciferase reporter, with and without adenoviral transduction of DNp38, were stimulated with cholecystokinin (CCK) or tumor necrosis factor-alpha (TNF-alpha) prior to measuring NF-kappaB activation.
CCK- or TNF-alpha-stimulated NF-kappaB-dependent gene transcription (luciferase assay) was substantially subdued by DNp38 expression. These findings were confirmed by electrophoretic mobility shift assay. Nuclear translocation of the p65 NF-kappaB subunit following agonist stimulation was evident (supershift). Characterization studies showed excellent adenoviral infection efficiency and cell viability in our AR42J cell model. Agonist-stimulated dose- and time-dependent p38 activation, with inhibition by DNp38 expression, was also confirmed.
The p38 MAP kinase regulates NF-kappaB pathway activation in exocrine pancreatic cells, and thus potentially plays a role in the mechanism of acute pancreatitis pathogenesis..
丝裂原活化蛋白激酶（MAP）激酶 p38 在急性胰腺炎发病机制中的作用存在争议。我们假设 p38 在调节胰腺外分泌细胞中 NF-κB 的激活中起作用。
用含有 NF-κB 反应性荧光素酶报告基因的 AR42J 细胞，以及转导 DNp38 的腺病毒，用胆囊收缩素（CCK）或肿瘤坏死因子-α（TNF-α）刺激，然后测量 NF-κB 的激活。
CCK 或 TNF-α 刺激的 NF-κB 依赖性基因转录（荧光素酶测定）被 DNp38 表达显著抑制。这些发现通过电泳迁移率变动分析得到了证实。激动剂刺激后，p65 NF-κB 亚单位的核易位是明显的（超迁移）。特征研究表明，我们的 AR42J 细胞模型具有良好的腺病毒感染效率和细胞活力。激动剂刺激的 p38 激活具有剂量和时间依赖性，DNp38 表达抑制了这种激活。
丝裂原活化蛋白激酶 p38 调节胰腺外分泌细胞中 NF-κB 途径的激活，因此可能在急性胰腺炎发病机制的机制中起作用。
