Development of a novel therapeutic approach to the renal injury by the control of anaphylatoxins.

通过控制过敏毒素开发一种新的肾损伤治疗方法。

• 批准号: 13557085
• 负责人: MATSUO Seiichi
• 金额: $ 5.7万
• 依托单位: NAGOYA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13557085/
• 关键词: complement; anaphylatoxin; C5a; gene transfer; carboxypeptidase R (CPR); rat; glomerulonephritis; anti-GBM antibody; 腎障害; 血栓形成性糸球体腎炎; トロンボモジュリン

In the present study, we investigated the possibility that arginine specific carboxypeptidase (CPR) is used as the effective anti-naphylatoxin agent in vivo. It is well known that CPR inactivate anaphylatoxins such as C5a by removing the argine residue. In order to test this hypothesis, we first cloned the rat homolog of human pro-carboxypeptidase R (pro-CPR). We then evaluated the efficacy and the reliability of in vivo gene transfer technique by retrograde injection of human erythropoietin (EPO) gene via tail vein. The EPO gene was efficiently introduced into the liver cells by this method. Then, we used rat homolog of pro-CPR gene in order to increase the concentration of pro-CPR in plasma. We found that 2 to 4 folds increase of plasma pro-CPR concentration was achieved. Then we induced anaphylatoxin-dependent diseases in rats. Rats treated with small amount of LPS and anti-glomerular basement membrane antibody (anti-GBM) showed very severe glomerular lesions characterized by the thrombotic glomerular capillary inflammation. Rats died within 24 hours after induction of diseases when they are not treated with anti-C5a receptor antagonists or thrombomodulin. The effects of pro-CPR gene transfer to these rats were limited. These results suggest that the further study is needed, for example, by the gene transfer of active form of CPR.

在本研究中，我们研究了精氨酸特异性羧肽酶（CPR）作为体内有效抗萘毒素剂的可能性。众所周知，CPR 通过去除精氨酸残基来灭活过敏毒素，例如 C5a。为了验证这一假设，我们首先克隆了人羧肽酶原 R (pro-CPR) 的大鼠同源物。然后，我们评估了通过尾静脉逆行注射人促红细胞生成素（EPO）基因的体内基因转移技术的有效性和可靠性。通过该方法将EPO基因有效地导入肝细胞中。然后，我们使用pro-CPR基因的大鼠同源物来增加血浆中pro-CPR的浓度。我们发现血浆 CPR 前体浓度增加了 2 至 4 倍。然后我们在大鼠中诱导过敏毒素依赖性疾病。用少量LPS和抗肾小球基底膜抗体（抗GBM）治疗的大鼠表现出非常严重的肾小球病变，其特征是血栓性肾小球毛细血管炎症。当不使用抗C5a受体拮抗剂或血栓调节蛋白治疗时，大鼠在诱发疾病后24小时内死亡。 pro-CPR 基因转移对这些大鼠的影响是有限的。这些结果表明需要进一步研究，例如通过 CPR 活性形式的基因转移进行研究。

项目成果

Ito, Isao: "Effects of a new synthetic selectin blocker in an acute rat thrombotic glomerulonephritis"Am J Kidney Dis. 38. 265-273 (2001)

Ito, Isao：“新型合成选择素阻滞剂对急性大鼠血栓性肾小球肾炎的影响”Am J Kidney Dis。

Ikeguchi, Hiroshi: "Effects of soluble human thrombomodulin in expreimental glomerulonephritis"Kidney Int. 61. 490-501 (2001)

池口宏：“可溶性人血栓调节蛋白对实验性肾小球肾炎的影响”Kidney Int。

Chika Kondo: "The role of C5a in the development of thrombolic glomerulonephritis in rats"Clinical and Experimental Immunology. 124. 323-329 (2001)

Chika Kondo：“C5a 在大鼠血栓性肾小球肾炎发展中的作用”临床和实验免疫学。

Masashi Mizuno: "Membrane complement regulators protect against the development of type II collagen-induced arthritis in rats"Arthritis and Rheumatism. 44. 2425-2434 (2001)

Masashi Mizuno：“膜补体调节剂可防止大鼠发生 II 型胶原诱导的关节炎”关节炎和风湿病。

Mizuno M, Nishikawa K, O.B Spiller, B.P Morgan, Okada N, Okada H, Matsuo S: "Membrane complement regulators protect against the development of type II collagen-induced arthritis in rats"Arthritis and Rheumatism. 44. 2425-2434 (2001)

Mizuno M、Nishikawa K、O.B Spiller、B.P Morgan、Okada N、Okada H、Matsuo S：“膜补体调节剂可防止大鼠发生 II 型胶原诱导的关节炎”关节炎和风湿病。