Investigation of transplantation cell therapy for acute renal endothelial cell injury.

移植细胞治疗急性肾内皮细胞损伤的研究。

• 批准号: 14370315
• 负责人: NOIRI Eisei
• 金额: $ 9.73万
• 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: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370315/
• 关键词: Cell transplantation therapy; Endothelial cell transplantation; Thrombotic micro-angiopathy; HUVEC; Ischemic acute renal failure; Chronic interstitial renal disease; GFP rat; Green fluorescent (GFP) rat; ヒト臍帯静脈欠陥内皮細胞

The governmental resource for medicare is about 3000 million yen per year in Japan. The population of ESRD is 200 thousand and the expense for each patient is 5 million yen per year. Thus 100 million yen is currently spent for ESRD in each year. Given the fact that the entry to ESRD is predominantly increasing from type 2 DM nephropathy, it is necessary to develop the therapeutic approach controlling the progressive renal disease.In this project, the efficacy of endothelial transplantation to renal disease was firstly investigated. The thrombotic micro-angiopathy (TMA) was induced to athymic nude rats injecting anti glomerular endothelial antibody super selectively to renal artery. The endothelial injury is initiated immediately after the injection, while its recovery process partly starts 3-4 days later as the endothelial proliferation. Therefore, the supplementation of endothelial cells to the defective sites may increase the number of regenerating glomeruli, accelerate the recovery … More process, and preserve renal function. Three days after the initiation of TMA, human umbilical venous endothelial cells (HUVE) were arterially injected to animals via catheter thru left jugular artery. One week after the injection of HJVEC, blood sampling was performed and the intact kidney was removed. After 24 h, the increase of BUN was examined and found the statistically significant increase because of the TMA kidney. This increase was reduced when HUVEC was transplanted to TMAA kidney. When rat renal interlobular arterial endothelial cells were injected instead of HUVEC, this increase was further decreased. The injected cells were observed at glomerular tuft even one week after the transplantation. It was also confirmed by electron microscopy. The hemodynamic effect of endothelial transplantation was examined using intravital video CCD camera which efficiently elucidated the peritubular no flow phenomena in ischemia reperfusion injury (ref 12). The glomerular red blood cell flow, initially low flow, was increased several minutes after the endothelial transplantation. Therefore the efficacy of endothelial transplantation in TMA model is partly derived from the improvement of glomerular blood flow.In parallel to these experiments, we developed green fluorescent rats to investigate the efficacy of bone marrow transplantation to several renal disease model, such as TMA, ischemic acute renal failure, chronic interstitial renal disease, SHC-renal failure, and crescentic glomerular nephritis. These projects are currently ongoing. During this project, at least one group reported the efficacy of bone marrow transplantation to ischemic acute renal failure though we could find deterioration during this short period. Therefore, the excretion factors from bone marrow cells were considered rather pivotal instead of transplanted bone marrow cells. Among the factors investigated, G-CSF is one of the promising factors related to stem cell transplantation, that was effective for the recovery process of ischemia-reperfusion injury, and that mechanism is currently investigating. In addition to these cell transplantation experiments, we elucidated the efficacy of hydroxyl radical scavenger to ischemia-reperfusion injury and got the acceptance of publication recently. This project was successfully conducted and generated valuable observations during the research periods. Less

日本每年用于医疗保险的政府资源约为30亿日元。ESRD的人口为20万，每位患者每年的费用为500万日元。因此，目前每年用于ESRD的费用为1亿日元。由于2型糖尿病肾病患者越来越多地进入终末期肾病（ESRD），因此有必要开发控制进展性肾病的治疗方法。在无胸腺裸大鼠肾动脉注射抗肾小球内皮细胞抗体，建立血栓性微血管病（TMA）模型。内皮损伤在注射后立即开始，而其恢复过程部分在3-4天后开始，因为内皮细胞增殖。因此，在损伤部位补充内皮细胞，可增加再生肾小球的数量，加速恢复 ...更多信息 处理和保护肾功能。在TMA开始后三天，通过导管经左颈动脉将人脐静脉内皮细胞（HUVE）动脉注射到动物体内。注射HJVEC后一周，进行血液取样并取出完整的肾脏。24 h后，检查BUN的增加，发现由于TMA肾，BUN的增加具有统计学意义。当HUVEC移植到TMAA肾时，这种增加减少。当注射大鼠肾小叶间动脉内皮细胞而不是HUVEC时，这种增加进一步降低。移植后1周仍能在肾小球内观察到注射细胞。电子显微镜也证实了这一点。使用活体视频CCD摄像机检查内皮移植的血流动力学效应，该摄像机有效地阐明了缺血再灌注损伤中的管周无血流现象（参考文献12）。肾小球红细胞流量，最初低流量，增加内皮移植后几分钟。因此，血管内皮移植在血栓形成模型中的疗效部分来自于肾小球血流的改善。在这些实验的同时，我们培育了绿色荧光大鼠来研究骨髓移植对几种肾脏疾病模型的疗效，例如血栓形成、缺血性急性肾衰竭、慢性间质性肾病、SHC肾衰竭和新月体肾小球肾炎。这些项目目前正在进行中。在这个项目中，至少有一个小组报告了骨髓移植对缺血性急性肾功能衰竭的疗效，尽管我们可以在这段短时间内发现恶化。因此，从骨髓细胞的排泄因子被认为是相当关键的，而不是移植的骨髓细胞。在所研究的因素中，G-CSF是干细胞移植相关的有希望的因素之一，其在缺血再灌注损伤的恢复过程中是有效的，并且其机制目前正在研究中。除了这些细胞移植实验外，我们还阐明了羟自由基清除剂对缺血再灌注损伤的作用，并于近期获得发表。该项目已成功实施，并在研究期间产生了宝贵的意见。少

项目成果

Yanase M, Ikeda H, Matsui A, Noiri E, Tomiya T, Arai M, Inoue Y, etc.: "HMG CoA reductase inhibitor modulates collagen GEL-contraction by hepatic myofibroblast-like stellate cell line : involvement of geranylgeranylated proteins."Com Hepatol. 14. S21 (200

Yanase M、Ikeda H、Matsui A、Noiri E、Tomiya T、Arai M、Inoue Y 等：“HMG CoA 还原酶抑制剂通过肝肌成纤维细胞样星状细胞系调节胶原蛋白 GEL 收缩：香叶基香叶基化蛋白的参与。”Com

Noiri E, Yamada S, Nakano A, Tsuchiya M, Masaki I, Fujino K, Nosaka K, Ozawa T, Fujita T, Uchida K: "Serum protein acrolein adducts : utilty in detecting oxidant stress in hemodialysis patients and reversal using a vitamin E-bonded hemodialyzer."Free Radi

Noiri E、Yamada S、Nakano A、Tsuchiya M、Masaki I、Fujino K、Nosaka K、Ozawa T、Fujita T、Uchida K：“血清蛋白丙烯醛加合物：检测血液透析患者氧化应激和使用维生素 E 逆转的实用性

Yanase M, Noiri E, et al.: "Functional diversity between Rho-kinase- and MLCK-mediated cytoskeletal actions in a myofibroblast-like hepatic stellate cell line."Biochem Biophys Res Commun. 305. 223-228 (2003)

Yanase M、Noiri E 等人：“肌成纤维细胞样肝星状细胞系中 Rho 激酶和 MLCK 介导的细胞骨架作用之间的功能多样性。”Biochem Biophys Res Commun。

Noiri E, Fujita T, Tokunaga K: "Endothelial nitric oxide synthase gene polymorphisms and renal survival"Hypertension. (in press). (2003)

Noiri E、Fujita T、Tokunaga K：“内皮一氧化氮合酶基因多态性与肾存活”高血压。

Noiri E, Fujita T, Tokunaga K: "Multifactorial Disease : Glu298asp of endothelial nitric oxide synthase"Hypertension. 41. e11-e12 (2003)

Noiri E、Fujita T、Tokunaga K：“多因素疾病：内皮一氧化氮合酶的 Glu298asp”高血压。