Neutrophils and NADPH oxidase mediate intrapancreatic trypsin activation in murine experimental acute pancreatitis
Neutrophils and NADPH oxidase mediate intrapancreatic trypsin activation in murine experimental acute pancreatitis
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DOI：
10.1053/gast.2002.32409
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发表时间：
2002-04-01
期刊：
影响因子：
29.4
通讯作者：
Pandol, SJ
中科院分区：
文献类型：
--
作者：
Gukovskaya, AS;Vaquero, E;Pandol, SJ
背景与目的：胰脏内消化酶的激活是胰腺炎实质细胞损伤的关键事件。我们假设，在胰腺炎期间募集到胰腺的中性粒细胞可能有助于这种激活。方法：采用大鼠、小鼠大剂量猪青蛋白治疗实验性胰腺炎。在胰腺匀浆中，采用荧光测定法和胰蛋白酶激活肽（TAP）抗体定位免疫细胞化学检测胰蛋白酶的激活。结果：与对照组相比，用抗中性粒细胞血清去除中性粒细胞的大鼠，高剂量蓝蛋白诱导的胰腺内胰蛋白酶激活和腺泡细胞TAP标记明显减弱。为了研究其机制，我们对缺乏烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶或髓过氧化物酶（MPO）的小鼠进行了胰蛋白酶激活研究。缺乏NADPH氧化酶的小鼠表现出蛋白诱导的胰蛋白酶活性的衰减，而缺乏MPO的小鼠则没有。通过Western blot分析、活性氧的产生和免疫细胞化学的测量，我们证明了NADPH氧化酶活性存在于中性粒细胞中，而不是胰腺腺泡组织中。结论：研究结果表明，浸润胰腺的中性粒细胞在急性胰腺炎患者消化酶的病理激活中发挥了新的作用，并表明这种作用是由NADPH氧化酶的产物介导的。
Background & Aims: Intrapancreatic activation of digestive enzymes is a key event in the parenchymal cell injury of pancreatitis. We hypothesized that neutrophils recruited to the pancreas during pancreatitis may contribute to such activation. Methods: To cause experimental pancreatitis, rats and mice were treated with high doses of cerulein. Activation of the digestive enzyme, trypsin, was measured in pancreatic homogenates using a fluorogenic assay and localized immunocytochemically with antibody to trypsin-activation peptide (TAP). Results: Compared with controls, rats depleted of neutrophils with antineutrophil serum exhibited a marked attenuation in intrapancreatic trypsin activation and acinar cell TAP labeling induced by high-dose cerulein. To examine the mechanism, mice deficient in either nicontinamide adenine dinucleotide phosphate (NADPH) oxidase, or myeloperoxidase (MPO) were studied for trypsin activation. Mice deficient in NADPH oxidase exhibited attenuation of the cerulein-induced trypsin activation, but those deficient in MPO did not. Using measurements of Western blot analysis, generation of reactive oxygen species, and immunocytochemistry, we demonstrated the NADPH oxidase activity is in neutrophils and not pancreatic acinar tissue. Conclusions: The results demonstrate a novel role for neutrophils infiltrating the pancreas in pathologic activation of digestive enzymes in acute pancreatitis and indicate that this effect is mediated by products of NADPH oxidase.