Transcriptional Regulation of the cardiovaocular genes in respone to stress

应激反应中心血管基因的转录调控

• 批准号: 11470156
• 负责人: KURABAYASHI Masahiko
• 金额: $ 6.59万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470156/
• 关键词: CARP; TRANSCRIPTION; VASCULAR SMOOTH MUSCLE CELL; TGF-β; PROMOTER; ANKYRIN; SIGNALING; GENE EXPRESSION; ストレス応答; BTEB2; Hex; スタチン; エンドセリン; 血管; 転写; 心肥大; 心不全; SHR; イソプロテノール; CARP; TRANSCRIPTION; VASCULAR SMOOTH MUSCLE CELL; TGF-β; PROMOTER; ANKYRIN; SIGNALING; GENE EXPRESSION; ストレス応答; BTEB2; Hex; スタチン; エンドセリン; 血管; 転写; 心肥大; 心不全; SHR; イソプロテノール

Despite the pleiotropic effects of TGF-β on vascular smooth muscle cells (VSMCs), only few genes have been characterized as direct targets of TGF-β in VSMCs. CARP, cardiac ankyrin repeat protein, has been thought to be expressed exclusively in the heart. In this study we showed that CARP is expressed in the vasculature after balloon injury and in cultured VSMCs in response to TGF-β. Analysis of a half-life of the cytoplasmic CARP mRNA levels and the transient transfection of the CARP promoter/luciferasere gene indicate that the regulation of CARP expression is increased by TGF-β at the transcriptional level. Transfection of expression vectors encoding Smads significantly activated the CARP promoter/luciferase activity. Deletion analysis and site-specific mutagenesis of the CARP promoter indicate that TGF-β response element is localized to CAGA motif at -108 bp relative to the transcription start site. Electrophoretic mobility shift assays showed that the binding activity to the CAGA motif was increased in nuclear extracts of cultured VSMCs by TGF-β. Cells transfected with adenovirus vector expressing CARP showed a significant decrease in DNA synthesis. Overexpression of CARP enhanced the TGF-β-mediated inhibition of the DNA synthesis. These data indicate that CARP is a downstream target of TGF-β/Smad-signaling in VSMCs, and suggest a role of CARP in mediating the inhibitory effects of TGF-β on the proliferation of VSMCs.

尽管TGF-β对血管平滑肌细胞（VSMC）产生了多效作用，但只有很少的基因被视为VSMC中TGF-β的直接靶标。鲤鱼，心脏arnkyrin重复蛋白，被认为仅在心脏中表达。在这项研究中，我们表明鲤鱼在球囊损伤后的脉管系统和培养的VSMC中表达，以响应TGF-β。分析细胞质鲤鱼mRNA水平的半衰期以及鲤鱼启动子/荧光素基因基因的瞬时翻译指标，表明TGF-β在转录水平上增加了鲤鱼表达的调节。编码Smads的表达载体的转染显着激活了鲤鱼启动子/荧光素酶活性。鲤鱼启动子的缺失分析和位点特异性诱变表明，相对于转录起始位点，TGF-β响应元件定位于-108 bp的CAGA基序。电泳迁移率偏移刺激表明，TGF-β培养的VSMC核提取物中与CAGA基序的结合活性增加。用表达鲤鱼的腺病毒载体翻译的细胞显示DNA合成显着降低。鲤鱼的过表达增强了TGF-β介导的DNA合成的抑制作用。这些数据表明CARP是VSMC中TGF-β/Smad信号的下游靶标，并表明鲤鱼在介导TGF-β对VSMC增殖的抑制作用中的作用。