Identification of the Receptor for ORAIP That Mediates Cardiac Response to Oxidative Stresses and Development of Treatment

介导心脏对氧化应激反应的 ORAIP 受体的鉴定和治疗方法的开发

• 批准号: 15390240
• 负责人: SEKO Yoshinori
• 金额: $ 8.32万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 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-15390240/
• 关键词: oxidative stress; cardiac myocytes; bioactive substance; autocrine; receptor; apoptosis; ischemia; reperfusion; hypoxia; reoxygenation; 酸化ストレス; 心筋細胞; オートクライン; アポトーシス; MAPK; 生理活性因子; 受容体

1.Production of recombinant ORAIP, a bioactive protein that mediates cardiac response to oxidative stresses :We subcloned a DNA sequence consisting of ORAIP cDNA+FLAG+(6X His) into an expression vector, then transfected the cDNA clone into an appropriate cell line. We purified recombinant ORAIP from the cell lysates with Ni column and an affinity column for FLAG protein.2.Identification of the ORAIP receptor :We collected the plasma membrane fraction from cultured cardiac myocyte lysates and solubilized it, then immunoprecipitated ORAIP receptor with recombinant ORAIP. Hypoxia/reoxygenation significantly increased the plasma membrane content of ORAIP receptor within 60 min of reoxygenation. Myocardial ischemia/reperfusion also significantly increased the expression of ORAIP receptor as well as ORAIP on the plasma membrane of cardiac myocytes within 30 min of reperftision.3.Identification of the cytoprotective factor that mediates cardiac response to oxidative stresses :We identified cyclophilin A (CyPA) as one of bioactive proteins released from cardiac myocytes in response to oxidative stresses. CyPA activated MAPK family members and Akt as well as increased Bcl-2 in cardiac myocytes. CyPA also increased Bcl-2 and Bcl-XL in cardiac fibroblasts. Taken together, CyPA seems to play a role in cardiac remodeling after ischemia/reperfusion by facilitating the proliferation of cardiac fibroblasts.

1.介导心脏氧化应激反应的生物活性蛋白--重组ORAIP的制备：我们将ORAIP的DNA序列亚克隆到表达载体中，然后将其导入到合适的细胞系中。2.ORAIP受体的鉴定：从培养的心肌细胞裂解液中收集质膜部分并溶解，然后用重组ORAIP进行免疫沉淀。缺氧/复氧60min内，ORAIP受体的质膜含量显著增加。心肌缺血/再灌流后30min，心肌细胞质膜上ORAIP受体和ORAIP的表达也显著增加。3.细胞保护因子的鉴定：我们鉴定亲环素A(CyPA)是心肌细胞在氧化应激反应中释放的生物活性蛋白之一。CyPA可激活心肌细胞内MAPK家族成员和Akt，增加Bcl2的表达。CyPA还可增加心脏成纤维细胞中Bcl2和Bclxl的表达。综上所述，CyPA似乎通过促进心脏成纤维细胞的增殖而在缺血/再灌流后的心脏重构中发挥作用。

项目成果

Hypoxia followed by re oxygenation induces secretion of cyclophilin A (CyPA) from cultured rat cardiac myocytes.

缺氧后进行再氧合可诱导培养的大鼠心肌细胞分泌亲环蛋白 A (CyPA)。

• 发表时间: 2004
• 期刊: Biochem Biophys Res Commun 317
• 作者: Seko Y;Fujimura T;Taka H;et al.
• 通讯作者: et al.

Seko Y, Sugishita K, Sato O, et al.: "Expression of costimulatory molecules (4-1BBL and Fas) and MHC class I chain-related A (MICA) in aortic tissue with Takayasu's arteritis."J Vas Res. 41. 84-90 (2004)

Seko Y、Sugishita K、Sato O 等人：“高安动脉炎主动脉组织中共刺激分子（4-1BBL 和 Fas）和 MHC I 类链相关 A (MICA) 的表达。”J Vas Res。

Takayasu myocarditis mediated by cytotoxic T lymphocytes

• DOI: 10.2169/internalmedicine.44.256
• 发表时间: 2005-03-01
• 期刊: INTERNAL MEDICINE
• 影响因子: 1.2
• 作者: Takeda, N;Takahashi, T;Nagai, R
• 通讯作者: Nagai, R

Genetic background influences therapeutic effectiveness of VEGF

• DOI: 10.1016/j.bbrc.2003.08.134
• 发表时间: 2003-10-10
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Fukino, K;Sata, M;Nagai, R
• 通讯作者: Nagai, R

Seko Y, Fukuda S, Nagai R: "Serum levels of endostatin, vascular endothelial growth factor (VEGF), and hepatocyte growth factor (HGF) in patients with acute myocardial infarction undergoing early reperfusion therapy."Clin Sci. (in press).

Seko Y、Fukuda S、Nagai R：“接受早期再灌注治疗的急性心肌梗死患者的内皮抑素、血管内皮生长因子 (VEGF) 和肝细胞生长因子 (HGF) 的血清水平。”《临床科学》。