Heat-Shock Protein 70-Overexpressing Gastric Epithelial Cells Are Resistant to Indomethacin-Induced Apoptosis
Heat-Shock Protein 70-Overexpressing Gastric Epithelial Cells Are Resistant to Indomethacin-Induced Apoptosis
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DOI：
10.1159/000215352
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发表时间：
2009-01-01
期刊：
影响因子：
3.2
通讯作者：
Yoshikawa, Toshikazu
中科院分区：
文献类型：
--
作者：
Hirata, Ikuhiro;Naito, Yuji;Yoshikawa, Toshikazu
背景/目的：保护肠粘膜免受非甾体抗炎药的影响仍然是一个尚未解决的问题。研究表明，线粒体损伤导致的上皮细胞凋亡是消炎痛所致胃粘膜损伤的重要发病机制。在这项研究中，我们揭示了过表达的热休克蛋白70（HSP 70）在吲哚美辛诱导的细胞凋亡和氧化应激的影响。研究方法：使用过表达HSP 70的大鼠胃粘膜细胞（7018-RGM-1细胞）和对照细胞（pBK-CMV-12细胞），并用0-500 μ M吲哚美辛处理24小时。分别通过WST-8测定法和乳酸脱氢酶释放测定法测量细胞活力和细胞毒性。Hoechst 33342和碘化丙啶荧光染色观察细胞凋亡。Bcl-2家族蛋白的表达，caspase-3的活化，和4-羟基-2-壬烯醛（4-HNE）修饰的蛋白质通过Western印迹分析进行了评估。结果：吲哚美辛可引起胃上皮细胞凋亡。与对照细胞相比，7018-RGM-1细胞在吲哚美辛处理后显著存活。在7018-RGM-1细胞中，促凋亡Bad蛋白的增加、抗凋亡Bcl-2蛋白的减少和半胱天冬酶的活化均被抑制。与对照细胞相比，在7018-RGM-1细胞中检测到较低水平的吲哚美辛诱导的4-HNE修饰。结论：过表达HSP 70可增强吲哚美辛诱导的胃上皮细胞损伤对凋亡和氧化应激的抵抗作用。版权所有（C）2009 S. Karger AG，巴塞尔
Background/Aims: Protecting intestinal mucosa from nonsteroidal anti-inflammatory drugs is still an unsolved problem. It has been revealed that apoptosis in epithelial cells as a result of mitochondrial injury is an important pathogenesis in indomethacin-induced gastric mucosal injury. In this study, we revealed the effect of overexpressed heat-shock protein 70 (HSP70) in indomethacin-induced apoptosis and oxidative stress. Methods: HSP70-overexpressing rat gastric mucosal cells (7018-RGM-1 cells) and control cells (pBK-CMV-12 cells) were used and treated with 0-500 mu M of indomethacin for 24 h. Cell viability and cytotoxity were measured by a WST-8 assay and a lactate dehydrogenase release assay, respectively. Apoptosis was observed by fluorescence microscopy staining with Hoechst 33342 and propidium iodide. The expression of Bcl-2 family proteins, activation of caspase-3, and 4-hydroxy-2-nonenal (4-HNE)-modified proteins were assessed by Western blot analysis. Results: Indomethacin caused apoptosis of gastric epithelial cells. The 7018-RGM-1 cells survived significantly after indomethacin treatment compared to the control cells. The increase in proapoptotic Bad proteins, the decrease in anti-apoptotic Bcl-2 proteins, and caspase activation were all suppressed in the 7018-RGM-1 cells. A lower level of indomethacin-induced 4-HNE-modification was detected in the 7018-RGM-1 cells than in the control cells. Conclusion: Overexpressed HSP70 may potentiate resistance to apoptosis and oxidative stress in indomethacin-induced gastric epithelial cell injury. Copyright (C) 2009 S. Karger AG, Basel