狼疮性肾炎热毒炽盛证和脾肾阳虚证的蛋白组学研究

Study of Proteomics on Kidney of Lupus Nephritis with Excessive Heat-toxic and Yang Deficiency of Spleen and Kidney

目的:应用蛋白质组学技术研究热毒炽盛型和脾肾阳虚型狼疮性肾炎及正常人之间的蛋白质差异,寻找热毒炽盛型和脾肾阳虚型狼疮性肾炎的相关蛋白及生物标记物。方法:采集狼疮性肾炎患者与正常人的血清,应用双向凝胶电泳、荧光染色及质谱分析技术分析其血清蛋白质表达差异,并对差异蛋白质进行分析。结果:发现狼疮性肾炎患者热毒炽盛型与脾肾阳虚型比较表达的差异点有5个,热毒炽盛型高表达的蛋白质是Ig mu chain C和间α-胰蛋白酶抑制因子重链H4;脾肾阳虚型高表达的蛋白质是细胞角蛋白-10。正常人与狼疮性肾炎患者的差异点有4个,狼疮性肾炎高表达的蛋白质是α1-抗胰蛋白酶和α1-抗糜蛋白酶,正常人高表达的是二硫键异构酶A6蛋白。结论:正常人与狼疮性肾炎患者的血清蛋白质表达有差异,这些差异蛋白可能有助于揭示狼疮性肾炎的发病机制或作为其新的生物标记物,对狼疮性肾炎的诊断及治疗有重要意义;热毒炽盛型与脾肾阳虚型狼疮性肾炎患者的血清蛋白质表达有差异。提示,同一疾病不同中医证型间的蛋白质表达不同,利用蛋白质组学技术有望对中医证候的发生机制进行阐释。

Objective ： Using proteomics technology with excessive heat-toxic and yang deficiency of spleen to study protein differences between lupus nephritis patients and kidney and normal human, and try to find out proteins that are associated with lupus nephritis and relevant biomarkers. Methods： Collecting serum from patients with lu- pus nepjritis and normal human, analyze the serum protein expression difference using two-dimensional gel electro- phoresis ,silver staining and Mass spectrometry. Results： The excessive heat-toxin type of LN patients had 5 differ- ential expression spots in contrast to yang deficiency of spleen and kidney of LN patients. The highly expressing proteins of the excessive heat-toxin type of LN patients were Ig mu chain C and Inter-alpha-trypsin inhibitor heavy chain H4, while the higy expressing protein of yang deficiency of spleen and kidney type of LN patients was Kera- tin type 1 cytoskeletal 10. The LN patients had 4 differential expression spots in contrast to normal persons. The highly expressing protein of LN patients were Alpha-l-antitrypsin and Alpha-l-antichymotrypsin, and the highly expressing protein of normal persons was protein disulfide-isomerase A6. Conclusion：There are some differences of proteins expressing between normal persons and Lupus Nephritis patients. The different proteins may help to reveal the pathogenesis of lupus nephritis or as new biomarkers for lupus nephritis. It is important in diagnosis and treat- ment of lupus nephritis, and also there are some differences of proteins expressing between excessive heat-toxic and yang deficiency of spleen and kidney in Lupus Nephritis patients, we may use proteomics technology to reveal the mechanism of TCM.