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A search by micro two-dimensional electrophoresis of an early marker in a diabetic nephropathy (2005)

通过显微二维电泳搜索糖尿病肾病的早期标记物（2005）

基本信息

批准号：
15580280
负责人：
SAKAGUTI Kazuko
金额：
$ 2.18万
依托单位：
Azabu 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 2005
项目状态：
已结题

项目摘要

PURPOSE : We searched for early checkup marker of a diabetes kidney disease with a clinical animal as a model.RESULT : Micro two-dimensional electrophoresis method analyzed a chronic kidney failure cat and plasma (1μL) of a diabetes cat in non-denaturation condition bottom ( I type) and non-denaturation-denaturation condition bottom (II type) and I used CBB dyeing and performed change analysis of a protein spot (the following, a spot). With chronic kidney failure cat plasma, IgM and two spots of Pre-albumin increased. Furthermore, increase of two spots was observed with II type in a low molecular weight domain. One this spot house which increased was identified with RBP by Western blot (WB) law. With diabetes cat plasma, increase was observed at one spot, and decrease was observed at two spots. The spot that increased was identified with Haptoglobin by WB method, but the spot that decreased is non-identification.We used 10 times concentration urine (1μL) for 100 times concentration urine, renal insufficiency cat urine for diabetes cat urine and performed change analysis of a spot likewise. With diabetes and a protein image of renal insufficiency cat urine, I was able to observe decrease of hoptoglobin (Hp) and albumin (Alb), transferrin (TO, prealbumin (Pre-alb) and increase of a spot of beta2 microglobulin (β_2-M). Although it was observed, as for the renal insufficiency cat urine, as for the individual difference, increase of all spots was remarkable. On the other hand, as for the diabetes cat urine, as for Pre-alb, β_2-M, increase was slightly observed although Alb, increase of Tf were remarkable. As a result, in Alb and the spot of Tf, in Pre-alb and the spot of β_2-M, possibility as marker-of renal insufficiency was suggested for an index of diabetes.

目的：以临床动物为模型，寻找糖尿病肾病的早期诊断标志物。结果：用显微双向电泳法分析了慢性肾功能衰竭猫和糖尿病猫血浆(1μL)在非变性状态下(I型)和非变性-变性状态下(II型)的变化，并用CBB染色进行了蛋白斑点的变化分析。慢性肾功能衰竭猫血浆中IgM和两点前白蛋白升高。此外，在低分子量域中观察到II型增加了两个斑点。经Western印迹(WB)法鉴定为RBP。糖尿病猫血浆1点升高，2点下降。用WB法用结合珠蛋白对增加的斑点进行鉴定，对减少的斑点不识别，用10倍浓缩尿(1μL)对100倍浓缩尿、肾功能不全猫尿对糖尿病猫尿进行斑点变化分析。对于糖尿病和肾功能不全猫尿的蛋白图像，我能够观察到跳珠蛋白(HP)和白蛋白(Alb)、转铁蛋白(TO)、前白蛋白(Pre-Alb)的减少以及β2微球蛋白(β_2-M)的增加。虽然观察到，但对于肾功能不全的猫尿，至于个体差异，各点的增加都是显著的。糖尿病猫尿中，前白蛋白、β_2-M略有升高，但Tf显著升高。提示白蛋白、Tf、前白蛋白、β_2-M可作为糖尿病患者肾功能不全的标志物。