【作者】 陈小瑞；

【导师】 申竹芳；

【作者基本信息】 中国协和医科大学 ， 药理学， 2004， 博士

【摘要】 糖尿病肾脏并发症严重威胁糖尿病患者的生活质量。无论是1型还是2型糖尿病患者，即使进行常规控制血糖的治疗，仍然有40％的患者最终出现肾脏并发症。在发达国家，因糖尿病肾病需要透析或肾移植治疗的终末期肾病大约占所有患者的40％。因而研究和开发治疗糖尿病肾病的药物具有重要意义。临床前实验研究是新药发现和研究的重要一环，本研究建立了有关糖尿病肾病临床前研究的几个重要方法，考察了1型和2型糖尿病动物模型的肾脏并发症发生和发展过程。并在1型和2型糖尿病动物模型上考察了几个化合物改善糖尿病肾脏并发症的作用及其作用机制。 临床前动物实验研究糖尿病肾病和一些其它类型的肾病时，最重要的检测指标是尿液微量白蛋白排泄量。冈为它直接反应了肾脏滤过功能的损害程度。本研究澄清了国内一些科研工作者在大小鼠尿液收集和尿液微量白蛋白测定上的若干模糊认识。建立了竞争性ELISA测定大鼠和小鼠尿液白蛋白的方法，该方法特异性强、易于推广，没有放射性污染，可定量测定大鼠和小鼠尿液微量白蛋白。用这种测定方法检测了1型、2型糖尿病大鼠模型以及2型糖尿病小鼠模型KKA~y小鼠的尿液白蛋白排泄量，相对于正常大鼠，糖尿病模型大鼠和小鼠均有尿液白蛋白排泄量的明显升高，提示这些动物有糖尿病肾脏并发症的出现。 非酶糖化终末产物是近些年来在包括肾病并发症在内的糖尿病慢性并发症研究中的一个热点，被认为在糖尿病以及糖尿病并发症的发生发展中是一个极重要的致病因素。本研究围绕非酶糖化终末产物做了三个方面的工作。一是结合国内外利用竞争性ELISA方法测定血清非酶糖化终末产物的文献资料，在本实验室制备了非酶糖化终末产物，用制备的非酶糖化终末产物免疫兔获得多克隆抗非酶糖化终末产物抗血清，进而建立了ELISA测定血清非酶糖化终末产物的方法，考察了吐温-20在测定中的作用。用这种方法测定1型糖尿病大鼠模型血清非酶糖化终末产物浓度，结果显示1型糖尿病模型大鼠血清非酶糖化终末产物明显升更多还原

【Abstract】 About 40% diabetic patients develop kidney complications eventually. Diabetic nephropathy is a leading cause of end-stage renal failure, accounting for 35% to 40% of all new cases requiring dialysis therapy throughout the world. Preclinical study is very important for finding new drugs to treat, diabetic nephropathy. In this study several methods very useful to preclinical study of diabetic nephropathy were developed and the ameliorative effects of several agents on diabetic nephropathy in rat and mouse diabetic models were observed.Part I A competitive ELISA for albumin in rat urine (attached: a competitive ELISA for albumin in mouse urine)Nephropathy is characterized by urinary micro-albumin. Rat is usually used in experimental studies. But there was no specific and simple method for detecting rat urinary albumin. A specific, easily operated and sensitive method for quantitatively determining rat urinary albumin is needed. Using rabbit anti-rat albumin polyclonal antibody, a competitive ELISA for rat albumin in urine was developed. With this method the urinary albumin in diabetic and normal rats was detected. The rabbit anti-rat albumin polyclonal antibody showed no cross reaction with bovine, human serum albumin and rat IgG. Based on the cross-reaction of the rabbit anti-rat albumin antibody with mouse albumin, a competitive ELISA for mouse urinary albumin was also established. The urinary albumin markedly increased in diabetic rats and mice.Part II Development and application of the ELISA method for measuring更多还原