REGULATORY MECHANISMS OF TRANSCRIPTIONAL REGULATORY FACTOR C/EBPdelta ON CELLULAR DEVELOPMENT OR HYPERTROPHY IN VASCULAR SMOOTH MUSCLE CELLS.

转录调节因子 C/EBPdelta 对血管平滑肌细胞发育或肥大的调节机制。

• 批准号: 08457210
• 负责人: HIWADA Kunio
• 金额: $ 4.99万
• 依托单位: EHIME UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457210/
• 关键词: vascular smooth muscle cells; cellular development; 5'-flanking region; PDGF receptor; transcriptional factor; CCAAT-binding protein; C; EBP family; promoter activity; 5'-上流域; EBPファミリー; 血管平滑筋; 遺伝子発現; サイトカイン; 血管壁再構築

C/EBPdelta is well known to be one of major members in CCAAT/enhancer-binding proteins. However, roles of C/EBPdelta on cellular development or hypertrophy in vascular smooth muscle cells (VSMC) are still unclear. In the present study, we have demonstrated that C/EBPdelta is not expressed in normal tissues, and its mRNA expression is observed in VSMC derived from genetically hypertensive rat, SHR.In addition, C/EBPdelta gene expression in VSMC is markedly induced by treatment with IL-1beta in a dose- or time- dependent manner. Structural and functional studies of the rat C/EBPdelta gene revealed that a TATA-like sequence (TAGAAAA) was located at 31-bp upstream region of the transcriptional start site, and an upstream control element (UCE) spanning -235 through -82 was essential for basic transcriptional activity of the gene. In addition, we also investigated effects of UCE on heterologous gene promoters including smooth muscle alpha-actin gene promoter and SV40 promoter. Interestingly, … More UCE specifically activated the promoter efficiency of alpha-actin gene suggesting that C/EBPdelta gene may be positively regulated by UCE via a cell- or promoter-type specific manner. Furthermore, to investigate roles of C/EBPdelta on pathological changes in the vessel wall, we performed an immunohistochemical detection of C/EBPdelta and platelet-derived growth factor alpha-receptor (PDGFalphaR), which is mainly controlled by C/EBPdelta at a transcriptional level, using damaged vessel walls after balloon injury. While a marked induction of C/EBPdelta protein was seen in smooth muscle layr or adventitia at the early phase (1-2 days after injury), PDGFalphaR protein was highly expressed in the neointima at the established phase (2-3 weeks after injury). These results strongly indicate that C/EBPdelta is functionally involved in the cellular development or hypertrophy in VSMC.In the future, we are planing to make C/EBPdelta-transgenic rats, and they are supposed to be good models to know a new function of C/EBPdelta in vivo. Less

众所周知，C/EBPdelta 是 CCAAT/增强子结合蛋白的主要成员之一。然而，C/EBPdelta 对血管平滑肌细胞 (VSMC) 细胞发育或肥大的作用仍不清楚。在本研究中，我们证明C/EBPδ在正常组织中不表达，并且在遗传性高血压大鼠SHR的VSMC中观察到其mRNA表达。此外，IL-1β以剂量或时间依赖性方式显着诱导VSMC中的C/EBPδ基因表达。对大鼠 C/EBPdelta 基因的结构和功能研究表明，TATA 样序列 (TAGAAAA) 位于转录起始位点上游 31 bp 区域，跨越 -235 至 -82 的上游控制元件 (UCE) 对于该基因的基本转录活性至关重要。此外，我们还研究了UCE对异源基因启动子的影响，包括平滑肌α-肌动蛋白基因启动子和SV40启动子。有趣的是，UCE 特异性激活 α-肌动蛋白基因的启动子效率，表明 C/EBPdelta 基因可能通过细胞或启动子类型特异性方式受到 UCE 的正向调节。此外，为了研究C/EBPδ对血管壁病理变化的作用，我们利用球囊损伤后受损的血管壁，对C/EBPδ和血小板源性生长因子α受体（PDGFαR）进行了免疫组织化学检测，PDGFαR主要在转录水平上受C/EBPδ控制。虽然在早期阶段（损伤后 1-2 天）在平滑肌层或外膜中观察到 C/EBPδ 蛋白的显着诱导，但 PDGFαR 蛋白在建立阶段（损伤后 2-3 周）在新内膜中高表达。这些结果有力地表明C/EBPδ在功能上参与了VSMC的细胞发育或肥大。将来，我们计划制作C/EBPδ转基因大鼠，它们应该是了解C/EBPδ体内新功能的良好模型。较少的

项目成果

Fukuoka T, Kitami Y, Kohara K, Hiwada K: "Molecular structure and function of rat CCAAT-enhancer binding protein-delta gene promoter." Biochem Biophys Res Commun. 231・1. 30-36 (1997)

Fukuoka T、Kitami Y、Kohara K、Hiwada K：“大鼠 CCAAT 增强子结合蛋白 δ 基因启动子的分子结构和功能。”Biochem Biophys Res Commun 231·1。

福岡 富和, 北見 裕, 大蔵 隆文, 間口 元文, 伊賀 瀬道也, 小原 克彦, 日和田 邦男: "血管平滑筋細胞における核内転写制御因子C/EBPδ遺伝子の発現調節" 血圧. 4・3. 265-271 (1997)

Tomikazu Fukuoka、Yutaka Kitami、Takafumi Okura、Motofumi Maguchi、Michiya Iga、Katsuhiko Ohara、Kunio Hiwada：“血管平滑肌细胞中核转录因子 C/EBPδ 基因表达的调节” 血压 4・3 .265-271（ 1997）

Fukuoka T,Kitami Y,Okura T,Maguchi M,Igase M,Kohara K,Hiwada K.: "Gene regulation of the transcriptional nuclear factor, C/EBPdelta gene, in vascular smooth muscle cells." Blood Pressure (in Japanese). 4 (3). 265-271 (1997)

Fukuoka T，Kitami Y，Okura T，Maguchi M，Igase M，Kohara K，Hiwada K.：“血管平滑肌细胞中转录核因子 C/EBPdelta 基因的基因调控。”

Kitami Y, Fukuoka T, Okura T, Takata Y, Maguchi M, Igase M, Kohara K, Hiwada K: "Molecular structure and function of rat platelet-derived growth factor β-receptor gene promoter." J Hypertens. 16(in press). (1998)

Kitami Y、Fukuoka T、Okura T、Takata Y、Maguchi M、Igase M、Kohara K、Hiwada K：“大鼠血小板衍生生长因子 β 受体基因启动子的分子结构和功能（正在出版）。” ）（1998）。

Kitami Y, Fukuoka T, Hiwada K, Inagami T: "Differential gene expression of the platelet-derived growth factor α-receptor in vascular smooth muscle cells of genetically hypertensive rats:reoles of CCAAT-enhancer binding proteins." Circ Res. (in press). (19

Kitami Y、Fukuoka T、Hiwada K、Inagami T：“遗传性高血压大鼠血管平滑肌细胞中血小板衍生生长因子 α 受体的差异基因表达：CCAAT 增强子结合蛋白的关系（见 Circ Res.）” (19