A possible treatment strategy for cardiovascular remodeling by metabolic syndrome-The gene therapy for use of MCP-1 7ND mutated gene-

代谢综合征引起的心血管重塑的可能治疗策略-利用MCP-1 7ND突变基因的基因治疗-

• 批准号: 15590781
• 负责人: KUWAHARA Fumitaka
• 金额: $ 2.24万
• 依托单位: KURUME UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590781/
• 关键词: Heart Failure; Cardiac Remodeling; Cardiac Fibrosis; MCP-1; OLETF Rat; Gene of 7ND; 7ND遺伝子; 左室拡張不全; 心・血管リモデリング; 炎症細胞浸潤; 酸化ストレス; TGF-β

Production of the MCP-17ND:Fc chimera cDNA (7ND) and verification of its dominant-negative activit :At first, we use PCR method to create 7ND cDNA by conjugating the amino-terminal deleted mutant of human MCP-1 and the human immunoglobulin Fc chain. Recombinant 7ND protein was obtained by overexpressing 7ND cDNA in 293 cells and we verified the dominant-negative activity of the recombinant 7ND protein on the MCP-1-induced macrophage production of interferon-gamma. In WKY rats and SD rats, 7ND gene transfer expressed 7ND protein in the muscle and significant elevation of circulating 7ND levels was documented by Elisa, having significant musclular damage at the site of injection (up to 1 g plasmid/kg body weight). However, we could not detect significant expression in circulating 7ND levels in 7ND (1 g/kg)-treated OLEFT rats. One explanation is the difference of the model. Another one is the muscle damage at the transfected site. Because OLEFT rats grow substantial larger than WKY and SD … More rats, the net dose of plasmid solution got much larger, resulting in the myocyte lysis and necrosis. Thus, we are now trying to search more efficient vectors.Evaluation of cardiac remodeling at multiple risk factor model (OLETF Rat) :We used OLETF rats to investigate cardiovascular remodeling in diabetes and hyperlipidemia status. LETF rats were use as controls. In stead of 7ND gene therapy, we used a neutralizing antibody against MCP-1 (NAb) to block MCP-1 activity. NAb or control IgG was administered everyday to OLETF and LETF rats from 30 weeks-old. At 40 weeks of age, we evaluated cardiac hypertrophy, fibrosis and cardiac function. In control OLETF rats had been greater interstitial myocardial fibrosis, especially reparative cardiac fibrosis, and sclerotic change of major vessels, compared with control LETF rats. Preceding cardiac fibrosis, macrophage infiltration and fibroblast proliferation were observed in perivascular space in control OLETF rats. This phenomenon suggested that hyperglycemia is involved in both inflammation changes and tissue damage remodeling. NAb treatment failed to reduce macrophage accumulation and cardiac fibrosis in OLETF rat hearts. We speculated that the inhibitory effects of NAb did not last for 10 weeks Less

MCP-17 ND：Fc嵌合体cDNA（7 ND）的制备及其显性负性活性的验证：首先，我们利用PCR方法将人MCP-1氨基端缺失突变体与人免疫球蛋白Fc链连接，构建了7 ND cDNA。通过在293细胞中过表达7 ND cDNA获得重组7 ND蛋白，并且我们验证了重组7 ND蛋白对MCP-1诱导的巨噬细胞产生干扰素-γ的显性负活性。在WKY大鼠和SD大鼠中，7 ND基因转移在肌肉中表达7 ND蛋白，并且通过Elisa记录了循环7 ND水平的显著升高，在注射部位具有显著的肌肉损伤（高达1g质粒/kg体重）。然而，我们在7 ND（lg/kg）处理的OLEFT大鼠中的循环7 ND水平中未检测到显著表达。一种解释是模型的不同。另一个是转染部位的肌肉损伤。因为OLEFT大鼠比WKY和SD大鼠长得大得多 ...更多信息 大鼠，质粒溶液的净剂量明显增加，导致心肌细胞溶解坏死。因此，我们现在正试图寻找更有效的载体。多危险因素模型（OLETF Rat）心脏重构的评价：我们使用OLETF大鼠研究糖尿病和高脂血症状态下的心血管重构。LETF大鼠作为对照。我们用抗MCP-1的中和抗体（NAb）来阻断MCP-1的活性，而不是7 ND基因治疗。从30周龄开始，每天向OLETF和LETF大鼠施用NAb或对照IgG。在40周龄时，我们评估了心脏肥大、纤维化和心功能。与对照组相比，对照组OLETF大鼠心肌间质纤维化，尤其是修复性心肌纤维化和主要血管的硬化改变更严重。在对照OLETF大鼠的血管周围空间中观察到先前的心脏纤维化、巨噬细胞浸润和成纤维细胞增殖。这一现象表明，高血糖既参与了炎症变化，也参与了组织损伤重塑。NAb治疗未能减少OLETF大鼠心脏中的巨噬细胞积聚和心脏纤维化。我们推测NAb的抑制作用不会持续10周。

项目成果

Kuwahara F, Kai H, et al.: "Hypertensive Myocardial Fibrosis and Diastolic Dysfunction"Hypertension. 43. 1-7 (2004)

Kuwahara F、Kai H 等：“高血压性心肌纤维化和舒张功能障碍”高血压。

Tokuda K, Kai H, Kuwahara F et al.: "Pressure-independent effects of angiotensin II on hypertensive myocardial fibrosis"Hypertension. 43. 499-503 (2004)

Tokuda K、Kai H、Kuwahara F 等人：“血管紧张素 II 对高血压心肌纤维化的压力独立效应”高血压。

Hypertensive myocardial fibrosis and diastolic dysfunction -Another model of inflammatioin?-

高血压心肌纤维化和舒张功能障碍-另一种炎症模型？-

• 发表时间: 2004
• 期刊: Hypertension 43(4)
• 作者: Kuwahara F;Kai H;et al.
• 通讯作者: et al.

Tokuda K, Kai H, Kuwahara F et al.: "Sub-depressor dose of angiotensin tipe-1 receptor blocker inhibits t-ansforming grouth factor-beta-mediated privascular fibrosis in hypertensive rat hearts"Journal of Cardiovascular Pharmacology. 42 Suppl 1. S61-S65 (2

Tokuda K、Kai H、Kuwahara F 等人：“亚抑制剂剂量的血管紧张素 Tipe-1 受体阻滞剂抑制高血压大鼠心脏中 t 型生长因子-β 介导的血管前纤维化”《心血管药理学杂志》。

Pressure-Independent Effects of Angiotensin II on Hypertensive Myocardial Fibrosis

• DOI: 10.1161/01.hyp.0000111831.50834.93
• 发表时间: 2004-02
• 期刊: Hypertension: Journal of the American Heart Association
• 作者: K. Tokuda;H. Kai;F. Kuwahara;H. Yasukawa;N. Tahara;Hiroshi Kudo;Kiyoko Takemiya;M. Koga;Tomoka Yamamoto;T. Imaizumi