优势抑制 p38 丝裂原活化蛋白激酶表达抑制 AR42J 细胞 NF-κB 激活。
Dominant negative p38 mitogen-activated protein kinase expression inhibits NF-kappaB activation in AR42J cells.
机构信息
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.
出版信息
Pancreatology. 2010;10(2-3):119-28. doi: 10.1159/000290656. Epub 2010 May 7.
BACKGROUND
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.
METHODS
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.
RESULTS
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.
CONCLUSION
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..
背景
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。
方法
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。
结果
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。
结论
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。
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