Generation and Analysis of Smooth Muscle-Specific C/EBP-δ Transgenic Rats

平滑肌特异性 C/EBP-δ 转基因大鼠的产生和分析

• 批准号: 12470155
• 负责人: HIWADA Kunio
• 金额: $ 7.94万
• 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: 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/en/grant/KAKENHI-PROJECT-12470155/
• 关键词: platelet-derived growth factor α-receptor; vascular smooth muscle cells; CCAAT; enhancer-binding protein-δ; transgenic rats; gene expression; 血管平滑筋; 血管壁リモデリング; EBPファミリー; EBPδ過剰発現; トランスジェニックラット; 細胞増殖因子; SMα-アクチンプロモータ; 組織特異的遺伝子発現

Platelet-derived growth factor (PDGF) is thought to play a significant role in various models of vascular remodeling, particularly in the early process of vascular diseases. Its action is mediated by its specific receptor, PDGF receptor (PDGFR). PDGF α-receptor (PDGFaR) plays an important role in the growth and proliferation of vascular smooth muscle, cells (VSMCs), and its gene expression is thought to be regulated by several potential transcriptional nuclear factors. However, the detailed mechanisms of tissue-specific transactivation of the PDGFαR gene in VSMCs remain to be clarified. We have previously demonstrated that the rat PDGFαR gene contains an enhancer core sequence for CCAAT/enhancer-binding proteins (C/EBPs) in its promoter region, and have also suggested that C/EBP-δ is the principal factor involved in the induction of tissue-specific transcriptional activity of the PDGFαR gene in VSMCs. To explore the definitive roles of C/EBP-δ protein on PDGFαR gene transcription in VSMCs, we here developed C/EBP-δ transgenic rats using a chimeric fusion gene of the mouse smooth muscle α-actin promoter and an entire coding region of rat C/EBP-δ cDNA. This report describes the first successful targeted overexpression of C/EBP-δ capable of inducing PDGFαR gene transcription and modifying cell proliferative activity to PBGFs. Targeted overexpression ofC/EBP-δ evokes high levels of PDGFαR gene expression, susceptibility to VSMC growth, and proliferation of VSMCs to PDGFs. The results obtained herein show evidence of a new role and new functional significance of C/EBP-δ on VSMC growth via PDGFαR during the process of vascular remodeling and atherosclerosis.

血小板源性生长因子（PDGF）被认为在各种血管重塑模型中发挥重要作用，特别是在血管疾病的早期过程中。其作用由其特异性受体PDGF受体（PDGFR）介导。PDGF α受体（PDGFaR）在血管平滑肌细胞（VSMCs）的生长和增殖中起重要作用，其基因表达被认为受多种潜在的核转录因子调控。然而，PDGFαR基因在VSMC中的组织特异性反式激活的详细机制仍有待阐明。我们先前已经证明大鼠PDGFαR基因在其启动子区含有CCAAT/增强子结合蛋白（C/EBP）的增强子核心序列，并且还表明C/EBP-δ是参与VSMC中PDGFαR基因组织特异性转录活性诱导的主要因子。为探讨C/EBP-δ蛋白对血管平滑肌细胞PDGFαR基因转录的影响，本研究采用小鼠平滑肌α-actin启动子与大鼠C/EBP-δ cDNA全编码区的嵌合融合基因构建了C/EBP-δ转基因大鼠。本文报道了首次成功的靶向过表达C/EBP-δ，其能够诱导PDGFαR基因转录并改变细胞向PBGF的增殖活性。靶向过表达C/EBP-δ可引起PDGFαR基因高水平表达，对VSMC生长和VSMC向PDGF增殖的易感性。本研究结果表明，C/EBP-δ在血管重塑和动脉粥样硬化过程中通过PDGFαR对VSMC生长具有新的作用和新的功能意义。

项目成果

Nakamura M,Okura T,Kitami Y,Hiwada K: "Nuclear factor 1 is a negative regulator of Gadd153 gene expression in vascular smooth muscle cells."Hypertension. 37[part2](In press). (2001)

Nakamura M、Okura T、Kitami Y、Hiwada K：“核因子 1 是血管平滑肌细胞中 Gadd153 基因表达的负调节因子。”高血压。

Igase M., Okura T., Nakamura M., Takata Y., Kitami Y., Hiwada K.: "Role of GADD153 (growth arrest- and DNA damage-inducible gene 153) in vascular smooth muscle cell apoptosis"Clin Sci. 100. 275-281 (2001)

Igase M.、Okura T.、Nakamura M.、Takata Y.、Kitami Y.、Hiwada K.：“GADD153（生长停滞和 DNA 损伤诱导基因 153）在血管平滑肌细胞凋亡中的作用”Clin Sci。

Yasunori Takata: "Peroxisome proliferator-γ activation inhibits intrleukin-1 β-mediated platelet-derived growth factor α-receptor gene expression via CCAAT/enhancer-binding protein-δ in vascular smooth muscle cells"Journal of Biological Chemistry. 276. 12

Yasunori Takata：“过氧化物酶体增殖物-γ 激活通过血管平滑肌细胞中的 CCAAT/增强子结合蛋白-δ 抑制白细胞介素-1 β 介导的血小板衍生生长因子 α-受体基因表达”《生物化学杂志》276. 12。

Okura T., Nakamura M., Takata Y., Watanabe S., Kitami Y., Hiwada K.: "Troglitazone induces apoptosis via the p53 and Gadd45 pathway in vascular smooth muscle cells"Eur J Pharmacol. 407. 227-235 (2000)

Okura T.、Nakamura M.、Takata Y.、Watanabe S.、Kitami Y.、Hiwada K.：“曲格列酮通过血管平滑肌细胞中的 p53 和 Gadd45 途径诱导细胞凋亡”Eur J Pharmacol。

Nakamura M., Okura T., Kitami Y., Hiwada K.: "Nuclear factor 1 is a negative regulator of Gadd153 gene expression in vascular smooth muscle cells"Hypertension. 37 (part 2). 419-424 (2001)

Nakamura M.、Okura T.、Kitami Y.、Hiwada K.：“核因子 1 是血管平滑肌细胞中 Gadd153 基因表达的负调节因子”高血压。