益肾活血法干预阿霉素肾病大鼠蛋白质组学的实验研究

Experimental study on proteomics of rats with adriamycin-induced nephropathy by tonifying the kidney and promoting blood circulation

目的:应用蛋白质组学筛选阿霉素肾病(AN)大鼠血清中差异表达的蛋白质,旨在蛋白质分子水平探索原发性肾病综合征(PNS)的诊断标志物及益肾活血中药肾康灵治疗的作用靶点。方法:实验分为正常组(A组)、AN病模组(B组)、AN病模+泼尼松组(C组)、AN病模+肾康灵组(D组)、AN病模+泼尼松+肾康灵组(E组)。利用表面增强激光解析-电离飞行时间质谱(SELDI-TOF-MS)联合CM10蛋白质芯片技术检测各组大鼠血蛋白质组质谱的表达,采用PBS II-C型蛋白质芯片阅读机自动收集数据。结果:在质荷比值(M/Z)2 000-50 000Da范围内,P<0.05,质谱仪筛选出B组与A组比较的11种差异蛋白,其中4种差异蛋白在B组中高表达,7种蛋白低表达;蛋白峰9069、15005、15047Da在B、C、D、E组间比较有显著差异,经鉴定可能分别为Cortexin-1、Interleukin-17A、Podoplanin,其中蛋白峰15005、15047从B组→C组→D、E组→A组呈递减趋势,蛋白峰9069呈递增趋势。结论:SELDI-TOF-MS能有效分析阿霉素肾病大鼠血清蛋白质质谱,益肾活血中药肾康灵干预治疗后蛋白质质谱表达有显著差异。

Objective：To analyze the blood proteomic patterns of rats with adriamycin-induced nephropathy by blood proteomics,for discussing the biomarker of primary nephrotic syndrome and target point of medication.Methods：Rats were divided into normal rats（group A）,adriamycin-induced nephropathy rats（group B）,adriamycin-induced nephropathy rats and prednisone（group C）,adriamycin-induced nephropathy rats and prednisone and Shenkangling（clinical density）（group D）,adriamycin-induced nephropathy rats and prednisone and Shenkangling（double clinical density）（group E）.Surface enhanced laser desorption/ionization-time of flight-mass spectrometry（SELDI-TOF-MS） CM10 protein clips was used to detect the blood proteomic patterns of every rat,PBS II-C protein clips reading machine was applied to collect data automaticly.Results：About 30 proteins with differential expression were detected in the mass charge ratio range of 2 000-5 0000（P〈0.05）,there were 11 proteins between B group and A group,the relative intensities in B group were significantly higher in 4 peaks,3 proteins decreased progressively from B group→C group→D,E group→A group,they were 9069,15005,15047Da,Cortexin-1,Interleukin-17A,Podoplanin.Conclusions：The blood proteomic patterns of rats with adriamycin-induced nephropathy can be analyzed in effect by SELDI-TOF-MS,significantly changed by tonifying the kidney and promoting blood circulation.