Over-expression of 70-kDa heat shock protein confers protection against monochloramine-induced gastric mucosal cell injury
Over-expression of 70-kDa heat shock protein confers protection against monochloramine-induced gastric mucosal cell injury
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DOI：
10.1016/j.lfs.2006.01.013
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发表时间：
2006-06-13
期刊：
影响因子：
6.1
通讯作者：
Watanabe, Sumio
中科院分区：
文献类型：
--
作者：
Oyake, Jinko;Otaka, Michiro;Watanabe, Sumio
已知主要的热休克蛋白HSP 70通过其作为分子伴侣的功能参与对环境应激的细胞保护。一氯胺（NH 2Cl）是一种强效的细胞毒性氧化剂，由嗜酸性粒细胞衍生的次氯酸和幽门螺杆菌尿素酶诱导的氨产生。为探讨HSP 70对NH_2Cl诱导的胃粘膜细胞损伤的保护作用，以含人HSP 70基因的pBK-CMV（7018-RGM-1）稳定转染大鼠胃粘膜细胞（RGM-1）或单纯转染pBK-CMV（pBK-CMV-12）作为对照，用不同浓度的NH_2Cl处理这些细胞。MTT法检测细胞活力，乳酸脱氢酶（LDH）释放法检测细胞膜直接损伤。DNA片段化分析显示NH 2Cl对pBK-CMV-12细胞有明显的损伤作用，并呈浓度依赖性。HSP 70过表达细胞系（7018-RGM-1）与对照细胞系（pBK-CMV-12）相比，NH_2Cl诱导的胃细胞损伤明显减轻，细胞坏死和凋亡均明显减少，提示HSP 70过表达对NH_2Cl诱导的胃细胞损伤具有重要的保护作用。(c)2006年爱思唯尔公司All rights reserved.
The major heat shock protein, HSP70, is known to be involved in cytoprotection against environmental stresses mediated by their function as a molecular chaperone". Monochloramine (NH2Cl) is a potent cytotoxic oxidant generated by neutrophil-derived hypochlorous acid and Helicobacter pylori urease-induced ammonia. In this study, to evaluate the cytoprotective effect of HSP70 against NH2Cl-induced gastric mucosal cell injury, rat gastric mucosal cells (RGM-1) were stably transfected with pBK-CMV containing the human HSP70 gene (7018-RGM-1) or pBK-CMV alone (pBK-CMV-12) as control cells.These cells were treated with various concentrations of NH2Cl. Cell Viability was determined by MTT assay and the direct plasma membrane damage was analyzed by lactate dehydrogenase (LDH) release assay. Apoptosis was determined by DNA fragmentation analysis.NH2Cl caused injury to pBK-CMV-12 cells in a concentration-dependent manner. NH2Cl-induced gastric cell injury was significantly deminished in HSP70 over-expressing cell line (7018-RGM-1) both necrosis and apoptosis compared to the control cell line (pBK-CMV-12) transfected with CMV vector alone.These result suggest that overexpression of HSP70 plays an important role in protecting gastric cells against NH2Cl-induced injury. (c) 2006 Elsevier Inc. All rights reserved.