Helicobacter pylori and H2O2 increase AP endonuclease-1/redox factor-1 expression in human gastric epithelial cells
Helicobacter pylori and H2O2 increase AP endonuclease-1/redox factor-1 expression in human gastric epithelial cells
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DOI：
10.1053/j.gastro.2004.06.017
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发表时间：
2004-09-01
期刊：
影响因子：
29.4
通讯作者：
Crowe, SE
中科院分区：
文献类型：
--
作者：
Ding, SZ;O'Hara, AM;Crowe, SE
背景和目标：幽门螺杆菌感染导致胃粘膜炎症、活性氧簇积累和氧化性DNA损伤。脱嘌呤/脱嘧啶核酸内切酶-1（APE-1）/氧化还原因子-1（Ref-1）修复受损的DNA并还原性激活转录因子，包括激活蛋白-1。考虑到H. pylori产生活性氧，活性氧调节其他细胞类型APE-1/Ref-1，我们检测了H. pylori、氧化应激和抗氧化剂对人胃上皮细胞APE-1/Ref-1表达的影响。方法：人胃粘膜上皮细胞系或从粘膜活检标本中分离的细胞用H。pylori、空肠弯曲杆菌和/或H2 O2。Western blot或逆转录聚合酶链反应检测APE-1/Ref-1的表达，间接常规免疫荧光和共聚焦免疫荧光检测APE-1/Ref-1在细胞内的分布。用[S-35]蛋氨酸标记法检测新蛋白的合成。通过使用含有3个激活蛋白1结合位点的荧光素酶连接的报告基因构建体评估APE-1/Ref-1功能。结果：APE-1/Ref-1蛋白和mRNA在静息状态胃上皮细胞中表达。APE-1/Ref-1蛋白表达在H2 O2或活的cad毒力岛H. pylori感染，而cag致病岛阴性H. pylori或C.空肠。H.幽门螺杆菌或活性氧介导的APE-1/Ref-1表达增加涉及抗氧化剂抑制的从头蛋白质合成。H. pylori或H2 O2也可诱导APE-1/Ref-1在核内聚集，APE-1/Ref-1过表达可增加激活蛋白1结合活性。结论：H. pylori感染或活性氧增强APE-1/Ref-1蛋白合成和细胞核积聚，提示APE-1/Ref-1参与了H.幽门感染
Background & Aims: Helicobacter pylori infection causes inflammation, accumulation of reactive oxygen species, and oxidative DNA damage in the gastric mucosa. Apurinic/apyrimidinic endonuclease-1 (APE-1)/redox factor-1 (Ref-1) repairs damaged DNA and reductively activates transcription factors, including activator protein-1. Considering that H. pylori generate reactive oxygen species and that reactive oxygen species modulate APE-1/Ref-1 in other cell types, we examined the effect of H. pylori, oxidative stress, and antioxidants on APE-1/Ref-1 expression in human gastric epithelial cells. Methods: Human gastric epithelial cell lines or cells isolated from mucosal biopsy samples were stimulated with H. pylori, Campylobacter jejuni, and/or H2O2 in the presence or absence of antioxidants. APE-1/Ref-1 expression was assayed by Western blot or reverse-transcription polymerase chain reaction, and its cellular distribution was determined by using indirect conventional and confocal immunofluorescence. New protein synthesis was detected by [S-35]methionine labeling. APE-1/Ref-1 function was assessed by using a luciferase-linked reporter construct containing 3 activator protein 1 binding sites. Results: APE-1/Ref-1 protein and messenger RNA were detected in resting gastric epithelial cells. APE-1/Ref-1 protein expression was increased after stimulation with H2O2 or live cad pathogenicity island-bearing H. pylori, but not cag pathogenicity island-negative H. pylori or C. jejuni. H. pylori- or reactive oxygen species-mediated increases in APE-1/Ref-1 expression involved de novo protein synthesis that was inhibited by antioxidants. H. pylori or H2O2 also induced nuclear accumulation of APE-1/Ref-1, and overexpression of APE-1/Ref-1 increased activator protein 1 binding activity. Conclusions: The data show that H. pylori or reactive oxygen species enhance APE-1/Ref-1 protein synthesis and nuclear accumulation in human gastric epithelial cells and implicate APE-1/Ref-1 in the modulation of the pathogenesis of H. pylori infection.