Eupatilin with Heme Oxygenase-1-Inducing Ability Protects Cultured Feline Esophageal Epithelial Cells from Cell Damage Caused by Indomethacin(Pharmacology)
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概要
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We previously reported that eupatilin (5,7-dihydroxy-3,4,6-trimethoxyflavone) extracted from Artemisia asiaitica, augmented the cellular antioxidant defense capacity through induction of the antioxidant protein heme oxygenase-1 (HO-1), thereby protecting ileal smooth muscle cells from nonsteroidal anti-inflammatory drug (NSAID)-induced intestinal toxicity. In the present study, we used cultured feline esophageal epithelial cells (EEC) to investigate the ability of eupatilin to induce expression of HO-1 and to analyze its cytoprotective effect against indomethacin-induced damage, since NSAID users have a higher risk of esophageal ulcers or esophagitis than non-NSAID users. A culture of EEC from cat was prepared. The identity of the cultures was confirmed by immunocytochemistry using cytokeratin antibodies. Western blot analysis showed a concentration- and time-dependent expression of HO-1 in response to eupatilin. Phosphorylation of extracellular regulating protein kinase (ERKs) and Akt, and nuclear translocation of nuclear related factor 2 (Nrf2) were induced by 150μM eupatilin in a time-dependent manner. Eupatilin-induced HO-1 expression and Nrf2 were partly attenuated by MEK inhibitor PD98059 and almost completely by phosphatidyl-inactiol 3 kinase (PI3K) inhibitor LY294002, but not by c-Jun N-terminal kinase (JNK) inhibitor SP600125 or p38 mitogen activated protein kinase (MAPK) inhibitor SB202190. MTT assay showed that treatment with 2mM indomethacin for 2h decreased cell viability to about 41%. Pre-treatment of cells with eupatilin resulted in the dose-dependent inhibition of indomethacin-induced cell damage. We confirmed that ZnPP, an HO-1 inhibitor, repressed eupatilin-induced HO-1 activity and showed the protective effect of eupatilin against indomethacin-induced cell injury. The data suggested that HO-1 was partly responsible for the eupatilin-mediated protective action of esophageal epithelial cells against indomethacin via both ERKs and PI3K/Akt pathways as well as Nrf2 translocation.
- 公益社団法人日本薬学会の論文
- 2009-04-01
著者
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MIN Young
Department of Internal Medicine, University of Ulsan College of Medicine, Asan Medical Center
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Sohn Uy
Department Of Pharmacology College Of Pharmacy Chung Ang University
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Min Young
Department Of Internal Medicine University Of Ulsan College Of Medicine Asan Medical Center
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Oh Tae
Research Laboratory Dong-a Pharmaceutical Co. Ltd.
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Song Hyun
Department Of Pharmacology College Of Pharmacy Chung Ang University
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SHIN Chang
Research Laboratory, Dong-A Pharmaceutical Co., Ltd.
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PARK Eon
Department of Pathology, College of Medicine, Chung Ang University
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Song Hyun
Department Of Internal Medicine Jeju National University School Of Medicine
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Park Eon
Department Of Pathology College Of Medicine Chung Ang University
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Shin Chang
Research Laboratory Dong-a Pharmaceutical Co. Ltd.
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Min Young
Department Of Pharmacology College Of Pharmacy Chung Ang University
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Song Hyun
Department Of Cardiovascular Surgery Asan Medical Center University Of Ulsan College Of Medicine
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MIN Young
Department of Herb Industry, Jungwon University
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