Liu Xiaodan, Yang Gang, Fan Qiuling, Wang Lining
Department of Nephrology, First Affiliated Hospital, China Medical University, Shenyang, Liaoning, China (mainland).
Med Sci Monit. 2014 Dec 17;20:2705-13. doi: 10.12659/MSM.893078.
Diabetic nephropathy (DN) is a leading cause of end-stage renal disease. To search for glomerular proteins associated with early-stage DN, glomeruli of spontaneous type 2 diabetic KKAy mice were analyzed by 2-dimensional differential gel electrophoresis (2D-DIGE).
Glomeruli of 20-week spontaneous type 2 diabetic KKAy mice and age-matched C57BL/6 mice were isolated by kidney perfusion with magnetic beads. Proteomic profiles of glomeruli were investigated by using 2D-DIGE and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Western blot analysis was used to confirm the results of proteomics. Immunohistochemical and semi-quantitative analysis were used to confirm the differential expression of prohibitin and annexin A2 in glomeruli.
We identified 19 differentially expressed proteins - 17 proteins were significantly up-regulated and 2 proteins were significantly down-regulated in glomeruli of diabetic KKAy mice. Among them, prohibitin and annexin A2 were up-regulated and Western blot analysis validated the same result in proteomics. Immunohistochemical analysis also revealed up-regulation of prohibitin and annexin A2 in glomeruli of KKAy mice.
Our findings suggest that prohibitin and annexin A2 may be associated with early-stage DN. Further functional research might help to reveal the pathogenesis of DN.
糖尿病肾病（DN）是终末期肾病的主要病因。为了寻找与早期DN相关的肾小球蛋白，采用二维差异凝胶电泳（2D-DIGE）分析了自发性2型糖尿病KKAy小鼠的肾小球。
通过磁珠肾脏灌注法分离20周龄自发性2型糖尿病KKAy小鼠和年龄匹配的C57BL/6小鼠的肾小球。采用2D-DIGE和基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱法研究肾小球的蛋白质组学图谱。采用蛋白质印迹分析来确认蛋白质组学的结果。采用免疫组织化学和半定量分析来确认禁蛋白和膜联蛋白A2在肾小球中的差异表达。
我们鉴定出19种差异表达蛋白——糖尿病KKAy小鼠肾小球中有17种蛋白显著上调，2种蛋白显著下调。其中，禁蛋白和膜联蛋白A2上调，蛋白质印迹分析验证了蛋白质组学中的相同结果。免疫组织化学分析也显示KKAy小鼠肾小球中禁蛋白和膜联蛋白A2上调。
我们的研究结果表明，禁蛋白和膜联蛋白A2可能与早期DN有关。进一步的功能研究可能有助于揭示DN的发病机制。
