Molecular pathobiological Study on Endothelial Function Ocurring during Vascular Remodeling

血管重塑过程中内皮功能的分子病理生物学研究

• 批准号: 10307003
• 负责人: SUEISHI Katsuo
• 金额: $ 24.26万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10307003/
• 关键词: gene transfer vector; Sev; VEGF; FGF-2; MCP-1; angiogenesis; atherosclerosis; vascular remodeling; 遺伝子導入; Sevベクター; PTCA; TGFβ^1; 遺伝子治療; 血栓症; 血管新生病; 転写因子; デコイオリゴ

The major results obtained in the last year (from Apr., 2000 to Mar., 2001) were as follows :1. Investigation on biological properties of Sendai virus vector (SeV) : 1) the in vitro and in vivo efficiencies of transduction and expression of beta-gal., luciferase, ecNOS, VEGF, FGF-2 and other genes using SeV were almost the same as those by Adeno virus vector, and SeV-mediated gene transfer was characterized by the fact that its brief exposure to cells in vitro and in vivo, less than 10 min., was enough for the peak gene expression. 2) Intaluminal injection of reporter gene-SeV could label not only ECs but also medial SMCs, but the neointima ocurred in human varicose veins hampered the transfection of medial SMCs.2. Molecular mechanism of angiogenesis in ischemic tissues : 1) Effect of intramuscular injection of VEGF and FGF-2 genes-SeV on development of collateral vessels in ischemic limb mouse model : injection of FGF-2 gene could induce endogeneous VEGF overexpression, and markedly i … More mprove blood perfusion in the ischemic limb by enhanced angiogenesis. However, VEGF gene injection worsened blood perfusion in ischemic limb. 2) the final and target angiogenic molecule for accelerating the development of collateral vessels in ischemic tissues was VEGF, but the harmonized collaboration of other angiogenic factors including FGF-2 was necessary for developing effective collateral circulation.3. In rat pulmonary hypertension induced by monocrotaline, interstitial remodeling by function of infiltrating macrophages through MCP-1 was revealed to be most important in development of pulmonary hypertension, and intramuscular infection of 7ND MCP-1 gene was effective for its prevention.4. Pigmentary retinopathy was revealed to be an angiogenic disease by the development of animal models of retroretinal angiogenesis by overexpression of VEGF using SeV.5. Diffuse intimal thickening was revealed to be an atherosclerosis-prone lesion by systematically and morphometrically examining human arteries including aorta, coronary artery, basilar artery of brain and others. Less

过去一年（2000年4月至2001年3月）取得的主要成果如下：仙台病毒载体（SeV）生物学特性的研究：1)β -gal在体外和体内的转导和表达效率。SeV介导的基因转染与腺病毒载体几乎相同，并且SeV介导的基因转染的特点是在体外和体内短暂暴露于细胞，不到10分钟就足以达到基因表达峰值。2)腹腔注射报告基因- sev不仅可以标记内皮细胞，还可以标记内侧的SMCs，但人静脉曲张中出现的新生内膜阻碍了内侧SMCs的转染。1)肌内注射VEGF和FGF-2基因- sev对缺血肢体小鼠模型侧支血管发育的影响：注射FGF-2基因可诱导内源性VEGF过表达，并通过增强血管生成明显改善缺血肢体的血液灌注。而VEGF基因注射使缺血肢体血流灌注恶化。2)加速缺血组织侧支血管发育的最终和目标血管生成分子是VEGF，但需要包括FGF-2在内的其他血管生成因子的协调协作才能形成有效的侧支循环。在单芥碱诱导的大鼠肺动脉高压中，经MCP-1浸润的巨噬细胞的间质重塑在肺动脉高压的发生发展中起重要作用，肌内感染7ND MCP-1基因可有效预防肺动脉高压的发生。利用SeV.5建立过表达VEGF的视网膜后血管生成动物模型，揭示了色素视网膜病变是一种血管生成疾病。通过对人体主动脉、冠状动脉、颅底动脉等动脉进行系统形态学检查，发现弥漫性内膜增厚是一种易发生动脉粥样硬化的病变。少

项目成果

Kitamoto S, Egashira K, Kataoka C, Koyanagi M, Katoh M, Shimokawa H, Morishita R, Kaneda Y, Sueishi K, Takeshita A.: "Increased activity of nuclear factor-κB participates in cardiovascular remodeling induced by chronic inhibition of nitric oxide synthesis

Kitamoto S、Egashira K、Kataoka C、Koyanagi M、Katoh M、Shimokawa H、Morishita R、Kaneda Y、Sueishi K、Takeshita A.：“核因子-κB 活性增加参与一氧化氮慢性抑制诱导的心血管重塑合成

Masaki I, et al: "Recombinant Sendai virus-mediated gene transfer to vasculature : a new class of efficient gene transfer vector to the vascular system."FASEB J. (in press).

Masaki I 等人：“重组仙台病毒介导的基因转移到脉管系统：一类新的有效基因转移载体到血管系统。”FASEB J.（出版中）。

Sakamoto T, et al: "Target gene transfer of tissue plasminogen activator to cornea by electric pulse inhibits intracameral fibrin formation and corneal cloudiness."Hum Gene Ther. 10. 2551-2557 (1999)

Sakamoto T 等人：“通过电脉冲将组织纤溶酶原激活剂的目标基因转移至角膜，抑制前房内纤维蛋白形成和角膜混浊。”Hum Gene Ther。

中島 豊: "Plaqueと組織因子" 動脈硬化. 26(1). 33-36 (1998)

Yutaka Nakajima：“斑块和组织因子”动脉硬化26(1)。

Ishibashi H, Nakagwa K, Onimaru M, Castellanous EJ, Kaneda Y, Nakashima Y, Shirasuna K, Sueishi K.: "Sp1 decoy transfected to carcinoma cells suppresses the expression of vascular endothelial growth factor, transforming growth factor β1, and tissue factor

Ishibashi H、Nakagwa K、Onimaru M、Castellanous EJ、Kaneda Y、Nakashima Y、Shirasuna K、Sueishi K.：“转染癌细胞的 Sp1 诱饵可抑制血管内皮生长因子、转化生长因子 β1 和组织因子的表达