肾康灵对阿霉素肾病大鼠尿蛋白质组学的影响

Impact of Shenkangling( 肾康灵) on the Urinary Proteomics of Adriamycin Nephropathy Model Rats

目的探讨原发性肾病综合征的诊断标志物和中药肾康灵药物治疗靶点。方法将50只SD大鼠随机分为正常组、模型组、泼尼松组、中西药组、肾康灵组各10只。除正常组外其余各组大鼠于实验第1天按5.5 mg/kg剂量一次性尾静脉注射阿霉素建立阿霉素肾病大鼠模型。造模后1周,正常组和模型组上、下午各灌服蒸馏水3 ml;泼尼松组上午灌服泼尼松悬液5.5 mg/kg,下午经灌服蒸馏水3 ml;肾康灵组上午灌服肾康灵煎剂1.5 g/100 g,下午经胃灌服蒸馏水3 ml;中西药组上午灌服泼尼松悬液5.5 mg/kg,下午经胃灌服肾康灵煎剂1.5 g/100 g。各组均连续灌胃22天后收集大鼠尿液,利用i TRAQ定量分析技术联合质谱分析法鉴定各组差异蛋白。结果模型组与正常组大鼠的显著差异蛋白3个,其中高表达的蛋白是afamin,isoform CRA_a,低表达的蛋白是gro和Cystatin related protein 2。中西药组与模型组比较,显著差异表达的蛋白有6个,其中高表达的蛋白是apolipoprotein A-Ⅱ,Apolipoprotein A-Ⅳ,Vitronectin,osteopontin,Coagulation factorⅫ,低表达的蛋白是afamin,isoform CRA_a。肾康灵组与模型组比较,显著差异表达的蛋白有6个,其中高表达的蛋白是apolipoprotein A-Ⅱ,Complement factor B,Vitronectin,osteopontin,Coagulation factorⅫ,低表达的蛋白是afamin,isoform CRA_a。结论 gro、Cystatin related protein 2、afamin,isoform CRA_a可作为原发性肾病综合征的诊断标志物,afamin,isoform CRA_a、apolipoprotein A-Ⅱ、Complement factor B、Vitronectin,osteopontin、Coagulation factorⅫ可作为肾康灵的药物治疗靶点。

Objective To explore the diagnostic marker of primary nephrotic syndrome（ PNS） and the therapeutic target of Shenkangling（ 肾康灵） decoction. Methods Fifty SD rats were randomized into normal group,model group,prednisone group,the herb-western medicine group and the Shenkangling group,10 in each group. Except for the normal group rats in other groups were injected adriamycin 5. 5 mg / kg by tail vein on the first day of the test to establish adriamycin nephropathy rats models. One week after modeling,the normal group and the model group were given 3 ml distilled water in the morning and in the afternoon respectively by gavage. The prednisone group was given prednisone suspension 5. 5 mg / kg in the morning by gavage,3ml distilled water in the afternoon. The Shenkangling group was given Shenkangling decoction 1. 5 g /100 g in the morning by gavage,3ml distilled water in the afternoon. The herb-western medicine group was given prednisone suspension 5. 5 mg / kg in the morning by gavage,Shenkangling decoction 1. 5 g /100 g in the afternoon. After 22 days,urine was collected and differential proteins in all groups were examined by isobaric tags for relative and absolute quantitation（ i TRAQ） quantitative analysis technology combined with mass spectrometric method. Results There were three significantly differential proteins in the model group and the normal group,including highly expressed protein afamin,isoform CRA_ a,as well as lowly expressed proteins gro and Cystatin related protein 2. Compared the herb-western medicine group with the model group,there were six significantly differential proteins,including highly expressed proteins apolipoprotein A-Ⅱ,Apolipoprotein A-Ⅳ,Vitronectin,osteopontin and Coagulation factor Ⅻ,as well as lowly expressed protein afamin,isoform CRA_ a. Compared the Shenkangling group with the model group,there were six significantly differential proteins,including highly expressed proteins apolipoprotein A-Ⅱ,Complement factor B,Vitronectin,osteopontin and Coagulation factor Ⅻ,as well as lowly expressed protein afamin,isoform CRA_ a. Conclusion Gro,Cystatin related protein 2,afamin and isoform CRA_ a might be used as the diagnostic markers of PNS. Afamin,isoform CRA_ a,apolipoprotein A-Ⅱ,Complement factor B,Vitronectin,osteopontin and Coagulation factor Ⅻ might be used as the therapeutic target of Shenkangling.