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目的:研究人参皂甙Rbl对人肾小管上皮细胞表达蛋白质组的影响。方法:人肾小管上皮细胞株(HK-2)常规培养,随机分为两组,实验组加入5μg/ml的Rbl,对照组加入等量的培养基,药物作用20min,裂解细胞,提取全细胞蛋白。双向电泳(2-DE)分离,ImageMaster 2D Platinum v5.0软件进行差异表达蛋白质组分析,基质辅助激光解吸附离子化飞行时间质谱(MALDI-TOF-MS)鉴定蛋白质。结果:通过对2-DE图谱蛋白斑点的匹配及对比分析,实验组的2-DE图谱共检出蛋白斑点为429个,其中236个为表达增强的蛋白斑点;经质谱鉴定的与Rbl作用相关的4个差异表达的蛋白斑点包括:普通转录因子ⅡH亚基1变体、双链断裂修复蛋白rad-21同源物、富含亮氨酸重复序列蛋白-45、DNA依赖性蛋白激酶,它们均属于磷酸化蛋白质。结论:Rbl作用于人肾小管上皮细胞的表达蛋白质组与对照组相比较存在明显的差异;Rbl对人肾小管上皮细胞的作用极有可能是通过相应的细胞信号转导网络系统来实现的。
Objective: To study the effect of ginsenoside Rb1 on proteomics of human renal tubular epithelial cells. Methods: The human renal tubular epithelial cell line (HK-2) was cultured in a routine manner and randomly divided into two groups. The experimental group received 5μg / ml Rb1, while the control group received the same amount of medium. protein. Two-dimensional electrophoresis (2-DE) separation was performed with ImageMaster 2D Platinum v5.0 software for differential expression proteomic analysis and protein identification by matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF-MS). Results: By matching and analyzing 2-DE map protein spots, 429 protein spots were detected in the 2-DE map of the experimental group, of which 236 were protein spots with enhanced expression. The results of mass spectrometry were correlated with the effect of Rbl The four differentially expressed protein spots include the common transcription factor IIH subunit 1 variant, the double-strand break repair protein rad-21 homolog, the leucine-rich protein-45, the DNA-dependent protein kinase, All belong to phosphorylated proteins. Conclusion: The expression of Rb1 protein in human renal tubular epithelial cells is significantly different from that of the control group. The effect of Rb1 on human renal tubular epithelial cells is most likely achieved through the corresponding cellular signal transduction network system.