幽门螺杆菌诱导AGS胃上皮细胞中白细胞介素-8和环氧化酶-2的表达：由核因子-κB介导

Helicobacter pylori-induced expression of interleukin-8 and cyclooxygenase-2 in AGS gastric epithelial cells: mediation by nuclear factor-kappaB.

作者信息

Kim H, Lim J W, Kim K H

机构信息

Dept. of Pharmacology and Institute of Gastroenterology, Yonsei University College of Medicine, Seoul, Korea.

出版信息

Scand J Gastroenterol. 2001 Jul;36(7):706-16. doi: 10.1080/003655201300191969.

DOI:10.1080/003655201300191969

PMID:11444469

Abstract

BACKGROUND

Helicobacter pylori infection might activate nuclear factor-kappaB (NF-kappaB), a transcriptional regulator of inducible expression of inflammatory genes, interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2). We studied the role of NF-kappaB on expression of IL-8 and COX-2 in H. pylori-stimulated AGS gastric epithelial cells by using antisense oligonucleotide (AS ODN) for NF-kappaB subunit p50 and an antioxidant, glutathione (GSH) as well as a NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC).

METHODS

AGS cells were treated with p50 AS ODN, GSH or PDTC in the presence of H. pylori. mRNA expression and protein levels for IL-8 and COX-2 were determined by Northern blot analysis and Western blot analysis. Levels of IL-8, 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha) and thromboxane B2 (TXB2) were measured in the medium by enzyme-linked immunosorbent assay. NF-kappaB activation was examined by electrophoretic mobility shift assay.

RESULTS

H. pylori induced a time-dependent expression of mRNA and protein for IL-8 and COX-2 via activation of NF-kappaB and increased the levels of IL-8, 6-keto-PGF1alpha and TXB2, which were inhibited by GSH and PDTC. H. pylori-induced expression of IL-8 and COX-2 was blocked in AGS cells transfected with p50 AS ODN.

CONCLUSION

NF-kappaB may play a novel role in expression of IL-8 and COX-2 in H. pylori-induced gastric inflammation.

摘要

背景

幽门螺杆菌感染可能激活核因子-κB（NF-κB），它是炎症基因白细胞介素-8（IL-8）和环氧化酶-2（COX-2）诱导性表达的转录调节因子。我们通过使用针对NF-κB亚基p50的反义寡核苷酸（AS ODN）、一种抗氧化剂谷胱甘肽（GSH）以及一种NF-κB抑制剂吡咯烷二硫代氨基甲酸盐（PDTC），研究了NF-κB在幽门螺杆菌刺激的AGS胃上皮细胞中对IL-8和COX-2表达的作用。

方法

在存在幽门螺杆菌的情况下，用p50 AS ODN、GSH或PDTC处理AGS细胞。通过Northern印迹分析和Western印迹分析测定IL-8和COX-2的mRNA表达和蛋白质水平。通过酶联免疫吸附测定法测量培养基中IL-8、6-酮-前列腺素F1α（6-酮-PGF1α）和血栓素B2（TXB2）的水平。通过电泳迁移率变动分析检测NF-κB的激活情况。