Rebamipide attenuates nonsteroidal anti-inflammatory drugs (NSAID) induced lipid peroxidation by the manganese superoxide dismutase (MnSOD) overexpression in gastrointestinal epithelial cells.
Rebamipide attenuates nonsteroidal anti-inflammatory drugs (NSAID) induced lipid peroxidation by the manganese superoxide dismutase (MnSOD) overexpression in gastrointestinal epithelial cells.
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发表时间：
2012-04
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通讯作者：
Y. Nagano;H. Matsui;O. Shimokawa;A. Hirayama;M. Tamura;Y. Nakamura;T. Kaneko;K. Rai;H. Indo-H.-In
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作者：
Y. Nagano;H. Matsui;O. Shimokawa;A. Hirayama;M. Tamura;Y. Nakamura;T. Kaneko;K. Rai;H. Indo-H.-In
非类固醇抗炎药(NSAIDs)通常会导致胃溃疡和糜烂等胃肠道并发症。最近的发病机制研究表明，NSAIDs通过在线粒体产生超氧阴离子诱导胃上皮细胞发生脂质过氧化，而不依赖于环氧合酶抑制和随后的前列腺素缺乏症。尽管还不清楚，但非甾体抗炎药对线粒体氧化磷酸化或解偶联的损害与超氧阴离子的产生有关。在生理上，超氧化物在锰超氧化物歧化酶(MnSOD)的作用下，立即转化为过氧化氢和双原子氧。瑞巴匹特是一种抗溃疡药物，对非甾体抗炎药引起的胃肠道脂质过氧化有保护作用。我们推测瑞巴匹特可能通过增加线粒体MnSOD蛋白的表达和减少NSAID处理的胃和小肠上皮细胞的超氧阴离子渗漏来减轻脂质过氧化。首先，为了检测瑞巴匹特增加胃肠上皮细胞线粒体中MnSOD蛋白的表达，我们对胃RGM1细胞和小肠IEC6细胞进行了抗MnSOD抗体的Western blotting分析。其次，为了检测瑞巴匹特的预处理是否能减轻NSAID诱导的线粒体损伤和脂质过氧化，我们用非甾体抗炎药加或不加瑞巴匹特，并用特异的荧光指示剂检测。最后，为了验证瑞巴匹特的预处理是否能减少非甾体类抗炎药诱导的线粒体超氧阴离子泄漏，我们使用一种特定的自旋捕获试剂--细胞色素PMPO，用电子自旋共振(ESR)波谱检测了吲哚美辛处理的RGM1细胞的线粒体。瑞巴匹特可增加RGM1和IEC6细胞MnSOD蛋白的表达，减轻NSAID诱导的线粒体损伤和脂质过氧化作用。瑞巴匹特可显著降低线粒体超氧阴离子信号强度。结论：瑞巴匹特通过增加胃和小肠上皮细胞MnSOD蛋白的表达和减少线粒体超氧阴离子渗漏来减轻脂质过氧化作用。
Nonsteroidal anti-inflammatory drugs (NSAIDs) often cause gastrointestinal complications such as gastric ulcers and erosions. Recent studies on the pathogenesis have revealed that NSAIDs induce lipid peroxidation in gastric epithelial cells by generating superoxide anion in mitochondria, independently with cyclooxygenase-inhibition and the subsequent prostaglandin deficiency. Although not clearly elucidated, the impairment of mitochondrial oxidative phosphorylation, or uncoupling, by NSAIDs is associated with the generation of superoxide anion. Physiologically, superoxide is immediately transformed into hydrogen peroxide and diatomic oxygen with manganese superoxide dismutase (MnSOD). Rebamipide is an antiulcer agent that showed protective effects against NSAID-induced lipid peroxidation in gastrointestinal tracts. We hypothesized that rebamipide may attenuate lipid peroxidation by increasing the expression of MnSOD protein in mitochondria and decreasing the leakage of superoxide anion in NSAID-treated gastric and small intestinal epithelial cells. Firstly, to examine rebamipide increases the expression of MnSOD proteins in mitochondria of gastrointestinal epithelial cells, we underwent Western blotting analysis against anti-MnSOD antibody in gastric RGM1 cells and small intestinal IEC6 cells. Secondly, to examine whether the pretreatment of rebamipide decreases NSAID-induced mitochondrial impairment and lipid peroxidation, we treated these cells with NSAIDs with or without rebamipide pretreatment, and examined with specific fluorescent indicators. Finally, to examine whether pretreatment of rebamipide attenuates NSAID-induced superoxide anion leakage from mitochondria, we examined the mitochondria from indomethacin-treated RGM1 cells with electron spin resonance (ESR) spectroscopy using a specific spin-trapping reagent, CYPMPO. Rebamipide increased the expression of MnSOD protein, and attenuated NSAID-induced mitochondrial impairment and lipid peroxidation in RGM1 and IEC6 cells. The pretreatment of rebamipide significantly decreased the signal intensity of superoxide anion from the mitochondria. We conclude that rebamipide attenuates lipid peroxidation by increasing the expression of MnSOD protein and decreasing superoxide anion leakage from mitochondria in both gastric and small intestinal epithelial cells.