Proteome analysis of diabetic-related proteins in rat kidney by fluorogenic derivatization–liquid chromatography–tandem mass spectrometry (FD-LC-MS/MS) method

• Main Authors: Pei-Yun Tsai, 蔡佩芸
• Other Authors: 李仁愛
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/74321667674778863473

碩士 === 臺北醫學大學 === 藥學研究所 === 98 === Diabetic nephropathy (DN) is a complication of diabetes mellitus which may lead to end-stage renal disease. Throughout its early course, DN has no symptoms. The first laboratory abnormality is a positive sign by microalbuminuria test. However, at this stage, a kidney biopsy clearly shows DN. Therefore, this study is designed to establish a new method to identify the differential expression of proteins in the kidney of control and streptozotocin (STZ)-induced diabetic rats in order to discover specific indicators for early diagnosis of DN. The Spregue-Dawley rats were sacrificed at 1, 4, 12, 24 weeks after i.p. injection of sodium citrate buffer or STZ. A thiol-specific fluorogenic reagent, 4-[2-(dimethylamino)ethyl]-7-chloro-2,1,3-benzoxadiazole-4-sulfonamide (DAABD-Cl) was utilized for the derivatization of proteins and then the derivatives were quantified and collected by high performance lipid chromatography with fluorescence detection (FD-HPLC). The isolated derivatives will be further identified by liquid chromatography tandem mass spectrometry (LC-MS/MS) with the MASCOT database searching system. After 1 week of injection, the expressions of approximately 7 proteins significantly changed. Only one protein had significantly changed after 4 weeks of injection. However, after 12 weeks of treatment, the number of altered proteins rose to 9. In addition, the changes appeared before the increase of microalbumin in the urine. The results may serve to find new biomarkers to improve early detection and new drug development in diabetic-related nephropathy.