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目的：建立丹参作用后人肾小管上皮细胞蛋白质组双向电泳分离体系，提高其分辨率和重复性。方法：丹参作用于人肾小管上皮细胞，提取全蛋白，按标准条件对蛋白质进行双向电泳分离，并对各个关键因素进行了优化。结果：最终选择的裂解液配方是1%TBP，4%CHAPS，0.2%Bio-Lyte，40mmol/L Tris，8mol/L尿素，2mol/L硫脲；使用pH 4～7的IPG胶条，电泳参数根据Bio-Rad公司的预设方案进行调整；改良硝酸银法进行蛋白质斑点染色。从而获得了满意的蛋白质组双向电泳图谱。结论：成功建立了丹参作用后人肾小管上皮细胞蛋白质组双向电泳分离体系，具有较高的分辨率和重复性，为进一步较为系统、全面、准确的评价丹参及其复方制剂的作用机制和药效提供方法学基础。“,”Objective: This study aimed to establish a system of two-dimensional gel electrophoresis (2-DE) for separation of proteomes from Human Kidney Tubular Epithelial cells. Methods: After the cultured HK-2 cells were treated by salviae miltiorrhizae,the proteins were extracted, and separated by 2-DE in standard conditions with an optimization for each important experimental parameter. Results: The lysis buffer containing1%TBP, 4%CHAPS, 0.2%Bio-Lyte, 40mmol/L Tris, 8mol/L urea, 2mol/L thiourea was applied to extract proteins from cells. The passive rehydration scheme of samples was used to help the separation of large molecular weight proteins in the IPG gel. Electrophoresis parameters recommended by Bio-Rad Corporation were adjusted adequately. In the process of silver staining, a sufficient washing operation ensured a week background of 2-DE maps. Conclusions:Under above optimized conditions, 2-DE maps with high resolution have been reproducibly obtained. Therefore a 2-DE separation system for proteomes from HK-2 cells has been successful y established.