Molecular mechanism of apoptosis and differentiation in vascular smooth muscle cells: the role of c-Jun N-terminal kinase (JNK)

血管平滑肌细胞凋亡和分化的分子机制：c-Jun N端激酶（JNK）的作用

• 批准号: 12670673
• 负责人: AOKI Hiroki
• 金额: $ 2.18万
• 依托单位: Yamaguchi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670673/
• 关键词: atherosclerosis; phenotypic modulation; MAP kinase; JNK; vascular smooth muscle; oxidative stress; intracellular signaling; adenovirus

[ Aim ] Recent studies indicate that the phenotypic modulation and apoptosis of vascular smooth muscle are fundamental phenomena underlying atherosclerosis. Vascular smooth muscle cells change their phenotype from highly differentiated contractile phenotype that is characterized by high expression of contractile proteins to synthetic one that is characterized by low expression of contractile proteins and high rate of protein synthesis. On the other hand, apoptosis of vascular smooth muscle cells may cause the loss of smooth muscle cell layer form arterial wall and destabilization of atherosclerotic lesion. The aim of this project is to elucidate the role of mitogen-activated protein kinases (MAP kinases), especially c-Jun N-terminal kinase (JNK), in phenotypic modulation and apoptosis of vascular smooth muscle cells. [ Results ] We established highly differentiated vascular smooth muscle cell culture system as a model system. Platelet-derived growth factor (PDGF) that initiates atheros … More clerotic change preferentially activated ERK family of MAP kinases, whereas oxidative stress by hydrogen peroxide preferentially activated JNK, suggesting different MAP kinase subfamily are mediating intracellular signaling in response to different extracellular stimuli. Both PDGF and hydrogen peroxide caused dedifferentiation of vascular smooth muscle cells in culture. In addition, hydrogen peroxide also caused apoptotic cell death. We made adenoviral vectors encoding dominant negative and wild type JNK, dominant negative and constitutively active SEK1 and MKK7, the upstream regulator of JNK pathway. These adenoviral vectors successfully transduced almost 100 % of vascular smooth muscle cells in culture. Expression of active SEK1 caused dedifferentiation of vascular smooth muscle cells partly mimicking the effect of oxidative stress. We also found that JNK pathway plays a central role in apoptosis of adult cardiac myocytes in culture. We identified a group of genes that are regulated under the control of JNK pathway by transcriptome analyses using DNA microarray. Less

[目的]近年来的研究表明，血管平滑肌的表型转化和细胞凋亡是动脉粥样硬化的基本现象。血管平滑肌细胞的表型由以高表达收缩蛋白为特征的高分化收缩表型转变为以低表达收缩蛋白和高蛋白质合成速率为特征的合成表型。另一方面，血管平滑肌细胞的凋亡可能导致动脉壁中的平滑肌细胞层丢失，导致动脉粥样硬化病变的不稳定。本研究旨在阐明丝裂原活化蛋白激酶(MAP)，尤其是c-jun氨基末端激酶(JNK)在血管平滑肌细胞表型调控和细胞凋亡中的作用。[结果]建立了高分化血管平滑肌细胞培养体系作为模型体系。启动动脉粥样硬化…的血小板衍生生长因子更多的硬化性改变优先激活ERK家族的MAP激酶，而过氧化氢引起的氧化应激优先激活JNK，这表明不同的MAP激酶亚家族在不同的细胞外刺激下介导细胞内信号转导。PDGF和过氧化氢均可引起体外培养的血管平滑肌细胞去分化。此外，过氧化氢还导致细胞凋亡。我们构建了编码显性负性和野生型JNK、显性负性和结构性活性的SEK1和JNK途径上游调节因子MKK7的腺病毒载体。这些腺病毒载体几乎100%成功地转导了培养的血管平滑肌细胞。活化的SEK1的表达导致血管平滑肌细胞去分化，部分模拟了氧化应激的影响。我们还发现JNK通路在培养的成人心肌细胞的凋亡中起着中心作用。通过DNA微阵列的转录组分析，我们发现了一组受JNK途径调控的基因。较少

项目成果

Yoshimura,Koichi et al.: "c-Jun N-terminal kinase plays a critical role in the oxidative stress-induced apoptosis of adult cardiac myocyte in vitro"Japanese Circulation Journal. 65 (1-A). 162-162 (2001)

Yoshimura、Koichi 等人：“c-Jun N 末端激酶在体外氧化应激诱导的成体心肌细胞凋亡中发挥着关键作用”《日本循环杂志》。

藤井幸蔵: "The role of stress-activated kinases in phenotypic modulation of vascular smooth muscle cell."Japanese Circulation Journal. 65(1-A). 523-523 (2001)

Kozo Fujii：“应激激活激酶在血管平滑肌细胞表型调节中的作用。”日本循环杂志 65(1-A) 523-523 (2001)。

青木浩樹: "Direct Activation of Mitochondrial Death Machinery by c-Jun N-terminal Kinse"Circulation. 104(17). II-247-LL-247 (2001)

Hiroki Aoki：“c-Jun N 末端激酶直接激活线粒体死亡机制”104(17) (2001)。

Aoki,Hiroki et al.: "Direct activation of mitochondrial apoptosis macchinery by c-Jun N-terminal kinase in adult cardiac mybcytes"Journal of Biological Chemistry. 277 (12) (in press). (2002)

Aoki、Hiroki 等人：“成人心脏 mybcytes 中 c-Jun N 末端激酶直接激活线粒体凋亡机制”《生物化学杂志》。

Aoki,Hiroki et al.: "Direct activation of mitochondrial death machinery by c-Jun N-tenninal kinase"Circulation. 104(17). II-247-II-247 (2001)

Aoki、Hiroki 等人：“c-Jun N-末端激酶直接激活线粒体死亡机制”循环。