Investigation on the molecular mechanism of JNK-mediated apoptosis in cardiac myocytes

JNK介导心肌细胞凋亡的分子机制研究

• 批准号: 14370229
• 负责人: AOKI Hiroki
• 金额: $ 6.34万
• 依托单位: Yamaguchi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370229/
• 关键词: oxidative stress; cardiac myocyte; apoptosis; JNK; mitochondria; cytochrome c; caspase; signal transduction; カスパーゼ; c-Jun N-terminal kinase; アデノウイルス・ベクター

Although oxidative stress causes activation of c-Jun N-terminal kinase (JNK) and apoptosis in many cell types, how the JNK pathway is connected to the apoptosis pathway is unclear, The molecular mechanism of JNK-mediated apoptosis was investigated in adult rat cardiac myocytes in culture as a model system that is sensitive to oxidative stress. Oxidative stress caused JNK activation, cytochrome c release and apoptosis without new protein synthesis. Oxidative stress-induced apoptosis was abrogated by dominant negative SEK1-mediated inhibition of JNK pathway, whereas activation of JNK pathway by constitutively active SEKI was sufficient to cause apoptosis. Inhibition of caspase-9, an apical caspase in mitochondrial apoptosis pathway, suppressed oxidative stress-induced apoptosis, whereas inhibition of caspase-8 had no effect, indicating that both JNK pathway and mitochondrial apoptosis machinery are central to oxidative stress-induced apoptosis. Both JNK and SEKI localized on mitochondria where JNK was activated by oxidative stress. Furthermore, active JNK phosphorylated several mitochondrial proteins and caused the release of apoptogenic factors such as cytochrome c from isolated mitochondria in a cell-free assay. These findings indicate that the JNK pathway is a direct activator of mitochondrial death machinery without other cellular components and provide a molecular linkage from oxidative stress to the mitochondrial apoptosis machinery. In addition, JNK activates matrix metalloproteinases in vascular smooth muscle cells and macrophages, which regulates tissue remodeling of cardiovascular system

虽然氧化应激导致c-Jun N-末端激酶（JNK）的激活和许多细胞类型的凋亡，但JNK通路如何与凋亡通路连接尚不清楚。在培养的成年大鼠心肌细胞中研究JNK介导的凋亡的分子机制，作为对氧化应激敏感的模型系统。氧化应激导致JNK激活、细胞色素c释放和细胞凋亡，而没有新的蛋白质合成。氧化应激诱导的细胞凋亡被显性负性SEK 1介导的JNK通路抑制所消除，而组成性活性SEKI激活JNK通路足以引起细胞凋亡。抑制caspase-9，线粒体凋亡途径中的顶端caspase，抑制氧化应激诱导的细胞凋亡，而抑制caspase-8没有影响，表明JNK途径和线粒体凋亡机制是中央氧化应激诱导的细胞凋亡。JNK和SEKI都定位于线粒体上，在线粒体中JNK被氧化应激激活。此外，活性JNK磷酸化几种线粒体蛋白，并导致在无细胞测定中从分离的线粒体释放促凋亡因子如细胞色素c。这些发现表明JNK途径是线粒体死亡机制的直接激活剂，而没有其他细胞组分，并提供了从氧化应激到线粒体凋亡机制的分子联系。此外，JNK还能激活血管平滑肌细胞和巨噬细胞中的基质金属蛋白酶，从而调节心血管系统的组织重构

项目成果

吉村耕一: "c-Jun N-terminal kinase regulates matrix metalloproteinase-9 activity in human abdominal aortic aneurysm"Circulation J. 67-I. 242 (2003)

Koichi Yoshimura：“c-Jun N-末端激酶调节人腹主动脉瘤中的基质金属蛋白酶-9 活性”Circulation J. 67-I（2003）。

Aoki, H.: "Direct activation of mitochondrial apoptosis machinery by c-Jun N-terminal kinase in adult cardiac myocytes"J Biol Chem. 277(12). 10244-10250 (2002)

Aoki, H.：“成人心肌细胞中 c-Jun N 末端激酶直接激活线粒体凋亡机制”J Biol Chem。

Yoshimura, K.: "c-Jun N-terminal kinase is required for development of abdominal aortic aneurysm in vivo"Circulation J. 68-1. 164 (2004)

Yoshimura, K.：“c-Jun N 末端激酶是体内腹主动脉瘤发展所必需的”Circulation J. 68-1。

吉村耕一: "c-Jun N-terminal kinase (JNK) governs the pathological extracellular matrix metabolism in human abdominal aortic"Circulation. 108. IV-193 (2003)

Koichi Yoshimura：“c-Jun N 末端激酶 (JNK) 控制人腹主动脉的病理性细胞外基质代谢”循环 108。 IV-193 (2003)

吉村耕一: "c-Jun N-terminal kinase (JNK) governs the pathological extracellular matrix metabolism in human abdominal aortic aneurysm"Circulation. 108. IV-193 (2003)

Koichi Yoshimura：“c-Jun N 末端激酶 (JNK) 控制人腹主动脉瘤的病理性细胞外基质代谢”循环 108。IV-193 (2003)