Development of new gene therapy for treatment of atherosclerosis using anti-apoptotic genes

开发利用抗凋亡基因治疗动脉粥样硬化的新基因疗法

• 批准号: 12670678
• 负责人: SUGANO Masahiro
• 金额: $ 2.18万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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/grant/KAKENHI-PROJECT-12670678/
• 关键词: Tumor necrosis factor; Soluble TNF receptor 1; Apoptosis; Angiogenesis; Endothelial cells; Myocardial infarction; TNF-alphaレセプタ-1; 酸化LDL

1) Growth of HUVEC transfected with sTNFRI vector also increased significantly compared to those transfected with control vector. Accumulation of sTNFRI significantly increased in conditioned medium from HUVEC transfected with sTNFRI vector compared to those transfected with control vector. HUVEC transfected with sTNFRI vector not only increased sTNFRI but also prevented shedding of sTNFRI from TNFRI. The TNF-a-induced internucleosomic fragmentation was also significantly prevented in HUVEC transfected with sTNFRI vector compared to those transfected with control vector.2) The incubation of HUVEC with oxLDL increased LOX-1 mRNA levels and CPP32-like protease activity, and induced apoptosis. Preincubation of HUVEC with nifedipine before incubation with oxLDL significantly suppressed the increase in LOX-1 mRNA levels and CPP32-like protease activity, preventing (7)___ in a dose-dependent manner. These results suggest that nifedipine blocks the suicide pathway leading to the apoptosis of endothelial cells by decreasing LOX-1 mRNA levels and CPP32-like protease activity.3) Here we report that direct injection of a sTNFRI expression plasmid DNA to the myocardium reduces infarct size in experimental rat AMI. Treatment with sTNFRI expression plasmid DNA reduced the TNF-alpha bioactivity in the myocardium and the apoptosis of cardiomyocytes. These findings suggest that the anti-TNF-alpha therapy by sTNFRI can be a new strategy for treatment of AMI.

1)转染sTNFRI载体的HUVEC的生长也明显高于转染对照载体的HUVEC。与转染对照载体的HUVEC相比，转染sTNFRI载体的HUVEC的条件培养基中sTNFRI的积累显著增加。转染sTNFRI载体的HUVEC不仅能增加sTNFRI，而且能阻止sTNFRI从TNFRI上脱落。与对照组相比，转染sTNFRI载体的HUVEC可明显抑制TNF-α诱导的核小体断裂。2）oxLDL可诱导HUVEC LOX-1 mRNA水平和CPP 32样蛋白酶活性升高，并诱导凋亡。在与oxLDL孵育之前，用硝苯地平预孵育HUVEC，显著抑制LOX-1 mRNA水平和CPP 32样蛋白酶活性的增加，以剂量依赖性方式阻止（7）_。这些结果表明硝苯地平通过降低LOX-1 mRNA水平和CPP 32样蛋白酶活性阻断了导致内皮细胞凋亡的自杀途径。3）在此我们报道了直接注射sTNFRI表达质粒DNA到实验性大鼠AMI的心肌中可以减少梗死面积。用sTNFRI表达质粒DNA处理降低了心肌中TNF-α的生物活性和心肌细胞的凋亡。提示sTNFRI抗TNF-α治疗AMI可能成为一种新的治疗策略。

项目成果

Masutomo K., Makino N., Sugano M., Fushiki S.: "Effects of Losartan on the Collagen Degradative Enzymes in Hypertrophic and Congestive types of Cardiomyopathic Hamsters"Mol Cell Biochem.. 224. 19-27 (2001)

Masutomo K.、Makino N.、Sugano M.、Fushiki S.：“氯沙坦对肥厚型和充血型心肌病仓鼠胶原蛋白降解酶的影响”Mol Cell Biochem.. 224. 19-27 (2001)

Hiroyoshi Hirayama, Masahiro Sugano, Nobuyuki Abe, Hidetoshi Yonemochi, Naoki Makino: "Troglitazone, an antidiabetic drug, improves left ventricular mass and diastolic funciton in normotensive diabetic patients"Int J. Cardiol. 77. 75-79 (2001)

Hiroyoshi Hirayama、Masahiro Sugano、Nobuyuki Abe、Hidetoshi Yonemochi、Naoki Makino：“曲格列酮是一种抗糖尿病药物，可改善血压正常的糖尿病患者的左心室质量和舒张功能”Int J. Cardiol。

Masahiro Sugano, Keiko Tsuchida, Hideharu Tomita, Naoki Makino: "Increased proliferation of endothelial cells with overexpression of soluble TNF-a receptor l gene"Atherosclerosis. (in press).

Masahiro Sugano、Keiko Tsuchida、Hideharu Tomita、Naoki Makino：“可溶性 TNF-α 受体 l 基因过度表达增加内皮细胞增殖”动脉粥样硬化。

Hiroyoshi Hirayama,Masahiro Sugano,Nobuyuki Abe,Hidetoshi Yonemochi,Naoki Makino.: "Troglitazone, an antidiabetic drug, improves left ventricular mass and diastolic function in normotensive diabetic patients."Int J.Cardiol.. 77. 75-79 (2001)

Hiroyoshi Hirayyama、Masahiro Sugano、Nobuyuki Abe、Hidetoshi Yonemochi、Naoki Makino.：“曲格列酮是一种抗糖尿病药物，可改善血压正常的糖尿病患者的左心室质量和舒张功能。”Int J.Cardiol.. 77. 75-79 (2001)

Masahiro Sugano, Keiko Tsuchida, Naoki Makino: "High-density lipoproteins protect endothelial cells from tumor necrosis factor-aloha -induced apoptosis"Biochem Biophys Res Comm.. 272. 872-876 (2000)

Masahiro Sugano、Keiko Tsuchida、Naoki Makino：“高密度脂蛋白保护内皮细胞免受肿瘤坏死因子-aloha 诱导的细胞凋亡”Biochem Biophys Res Comm.. 272. 872-876 (2000)