TRANSCRIPTIONAL REGULATION OF STRESS-RELATED GENES IN GASTRIC EPITHELIAL CELLS.

胃上皮细胞中应激相关基因的转录调控。

• 批准号: 07670596
• 负责人: ROKUTAN Kazuhito
• 金额: $ 1.66万
• 依托单位: SCHOOL OF MEDICINE,THE UNIVERSITY OF TOKUSHIMA
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670596/
• 关键词: GASTRIC EPITHELIAL CELLS; OXIDATIVE STRESS; HEAT SHOCK PROTEIN; TRANSCRIPTION FACTOR; HSF1; NF-kB; STRESS-RELATED GENES; 培養胃粘膜細胞; ストレス蛋白質遺伝子; グルタチオン

The first experiment was designed to reveal the novel action of an antiulcer drug, geranylgeranylacetone (GGA), which rapidly induces resistance of gastric mucosal cells to irritants in vivo and in vitro. GGA induced resistance of cultured guinea pig gastric mucosal cells against ethanol-induced exfoliation and damage within 30 min, proportionally to the induction of the HSPs. This protection was blocked by cycloheximide but not by indomethacin. GGA rapidly activated HSF1 and caused expression of HSP70 mRNA,suggesting that GGA may induce transcriptional activation of HSP genes. Intragastric administration of GGA to rats induced HSPs in gastric mucosa, and additionally augmented the expression of HSP70 mRNA and induction of HSPs in the mucosa of rats confronted with restraint and water-immersion stress. This novel action may increase gastric mucosal defense and reduce damage at times of stress conditions.Nuclear factor-kappa B (NF-kB) /Rel transcription factors play an important role in … More the transcriptional activation of a variety of genes involved in inflammatory and immune responses. The aims of second experiment were to identify the oxidant-sensitive components of NF-kB in gastric epithelial cells and to reveal the roles of this factor under oxidative stress. Gel mobility shift assay with a kB oligonucleotide showed that exposure of primary cultures of guinea pig gastric epithelial cells to hydrogen peroxide and diamide rapidly produced a DNA-protein complex within 5 min. The kB-binding proteins consisted of a p50/p65 heterodimer, which was identified by supershift experiments, UV-crosslinking analysis, immunoprecipitation, and immunocytochemical analyzes with antibodies against p50, p65, and c-Rel. Both hydrogen peroxide and diamide caused the up-requlation of p105 (a p50 precursor) synthesis and the expression of inducible nitric oxide mRNA,which have been suggested to be putative target genes for NF-kB.Thus, our results suggest that the oxidant-sensitive p50/p65 heterodimer may play an important role in the initiation of the responses of gastric epithelial cells to oxidative stress. Less

第一个实验旨在揭示抗溃疡药物香叶基香叶基丙酮（GGA）的新作用，它能快速诱导胃粘膜细胞对体内和体外的刺激物产生抵抗力，GGA诱导培养的豚鼠胃粘膜细胞对乙醇诱导的抵抗力。 30 分钟内剥落和损伤，与 HSP 的诱导成比例。这种保护被放线菌酮阻断，但不能被放线菌酮阻断。 GGA 快速激活 HSF1 并引起 HSP70 mRNA 的表达，表明 GGA 可以诱导 HSP 基因的转录激活，诱导胃粘膜中的 HSPs，并且还增强了 HSP70 mRNA 的表达并诱导了 HSPs。面对束缚和水浸应激的大鼠粘膜，这种新的作用可能会增强胃粘膜的防御能力并减少应激时的损伤。核因子-kappa B (NF-kB) /Rel 转录因子在参与炎症和免疫反应的多种基因的转录激活中发挥着重要作用。第二个实验的目的是鉴定对氧化剂敏感的基因。胃上皮细胞中 NF-kB 的成分，并揭示该因子在氧化应激下的作用，用 kB 寡核苷酸进行凝胶迁移率变化测定，结果表明豚鼠胃上皮原代培养物的暴露。细胞与过氧化氢和二酰胺在 5 分钟内迅速产生 DNA-蛋白质复合物，其中 kB 结合蛋白与 p50/p65 异二聚体连接，通过超位移实验、UV 交联分析、免疫沉淀和免疫细胞化学与抗体进行嵌套来鉴定。 p50、p65 和 c-Rel 均导致 p105（p50 前体）合成和诱导表达的上调。一氧化氮 mRNA，被认为是 NF-kB 的假定靶基因。因此，我们的结果表明，氧化剂敏感的 p50/p65 异二聚体可能在启动胃上皮细胞对氧化应激的反应中发挥重要作用。较少的

项目成果

Kutsumi,H.et al.: "Evidence for participation of vicinal dithiols in the activation sequence of the respiratoryburst of human neutrophils" Blood. 85. 2559-2569 (1995)

Kutsumi,H.等人：“邻位二硫醇参与人类中性粒细胞呼吸爆发激活序列的证据”血液。

Hiromu Kutsumi et al.: "Evidence for participation of vicinal dithiols in the activation sequence of the respiratory burst of human neutrophils." Blood. 85. 2559-2569 (1995)

Hiromu Kutsumi 等人：“邻位二硫醇参与人类中性粒细胞呼吸爆发激活序列的证据。”

Teshima,S.et al.: "Induction of heat shock proteins and their possible roles in macrophages during activation by macrophage colony-stimulating factor" Biochem.J.314. 497-504 (1996)

Teshima,S.et al.：“巨噬细胞集落刺激因子激活期间热休克蛋白的诱导及其在巨噬细胞中可能的作用”Biochem.J.314。

Keiichi Kawai et al.: "Kinetics of gastric epithelial cells in duodenal ulcer." J. Gastroenterol.30. 428-436 (1995)

Keiichi Kawai 等人：“十二指肠溃疡胃上皮细胞的动力学”。

Kutsumi, H., Kawai, K., Johnston, R.B.Jr., and Rokutan, K.: "Evidence for participation of vicinal dithiols in the activation sequence of the respiratory burst of human neutrophils" BLOOD. 85. 2559-2569 (1995)

Kutsumi, H.、Kawai, K.、Johnston, R.B.Jr. 和 Rokutan, K.：“邻位二硫醇参与人类中性粒细胞呼吸爆发激活序列的证据”BLOOD。