慢性心力衰竭患者尿液双向凝胶电泳技术的优化  

Optimization of two-dimensional polyacrylamide gel electrophoresis on urine samples from patients with chronic heart failure

作　　者：侯丽娜[1] 刘炜烨[1] 贾小平[1] 李亮[1] 李小溪[1] 

机构地区：[1]南方医科大学南方医院健康管理中心,广东广州510515

出　　处：《转化医学电子杂志》2017年第8期41-43,共3页E-Journal of Translational Medicine

基　　金：广东省科技厅科技计划项目(2016zc0078)

摘　　要：目的:分析慢性心力衰竭患者尿液双向电泳技术的影响因素以提高其分辨率及稳定性.方法:采用超滤法富集并浓缩尿液蛋白,Bradford法测浓度后进行双向电泳,凝胶经银染显色后,用Melanie 4软件分析双向电泳图谱.分别对蛋白上样量,聚焦参数等步骤进行优化.结果:超滤浓缩法,150μg的上样量,60 KVh的聚焦总伏特数,进行的双向电泳图谱分辨率高,稳定性好.结论:对上样量和聚焦总伏特数优化后可获得稳定性高和重复性好的慢性心力衰竭患者尿液双向电泳图谱,以便开展后续研究.AIM: To analyze the influence factors of two-dimensional electrophoresis( 2-DE) on urine samples from the patients with chronic heart failure and improve the resolution and stability of 2-DE map. METHODS: After using the method of ultrafiltration, the 2-DE was conducted after measuring the concentration of the total protein by Bradford method. The gel was scanned and analyzed by Melanie 4 following silver staining. We improved the sample volume of proteins,focus parameters and other related steps to optimize the 2-DE. RESULTS: We could successfully obtain a higher resolution and reproducible 2-DE image with ultrafiltration,protein loaded with 150 μg and focus voltage with a total of 60 KVh. CONCLUSION: The well-established 2-DE map on urine samples of chronic heart failure patients made by 2-DE is useful in studying the mechanism and treatments of chronic heart failure.

关 键 词：慢性心力衰竭 尿液 双向凝胶电泳 蛋白质组 

分 类 号：R541.61[医药卫生—心血管疾病]