环孢素肾毒性蛋白质标识物血浆双向电泳法建立的研究  

Establishment of Protocol of Two-dimensional Gel Electrophoresis to Identify Plasma Protein Biomarker of Cyclosporine Renal Toxicity

作　　者：舒展[1] 蒲小平[2] 李泉[2] 王悦[3] 翟所迪[1,4] 

机构地区：[1]北京大学第三医院药剂科 [2]北京大学药学院分子与细胞药理学系 [3]北京大学第三医院肾内科 [4]北京大学治疗药物监测和临床毒理中心,北京市100083

出　　处：《中国药房》2006年第12期899-901,共3页China Pharmacy

基　　金：教育部教育振兴行动计划特殊专项项目(北京大学"985"工程Ⅰ期项目)

摘　　要：目的建立环孢素肾毒性蛋白质标识物血浆双向电泳法。方法将大鼠随机分为低剂量组、高剂量组、空白对照组(蒸馏水),连续7d对低、高剂量组大鼠分别灌胃给予5、100mg/(kg.d)环孢素,qd。检查肾功能与尿—N—乙酰—β—氨基葡萄糖苷酶,观察肾病理变化,明确急性肾损伤的程度;取空白对照组和高剂量组大鼠的血浆样品,用硫酸铵分步盐析法去除其中的高丰度蛋白质后进行双向电泳与图谱对比。结果低剂量组未显示明显的肾损伤,高剂量组则成功地建立了环孢素急性肾损伤大鼠模型。经硫酸铵分步盐析处理后,空白对照组和高剂量组血浆样品的双向电泳图谱分辨率良好,并显示较为稳定的差异蛋白质斑点。结论建立的双向电泳法和技术体系可用于寻找环孢素急性肾毒性的血浆蛋白质标识物质。BJECTIVE： To establish a protocol of two - dimensional gel electrophoresis （2 - DE） in order to identify plasma protein marker of cyclosporine renal toxicity. METHODS： Wistar rats were randomly divided into control group （distilled water） , low dose group and high dose group. Rats in low dose group and high dose group were given cyclosporine at a dose of 5 mg/（kg · d） and 100 mg/（kg · d） by gavage, respectively. The extent of renal impairment was evaluated through biochemical assays of renal function and urease （N- acetyl-β- glucosaminidase） and pathological observation after a sevenday control. Rat plasma from control group and high dose group was sampled, and high abundance of protein was removed through stepwise precipitation with ammonium sulfate, then compared with each other by 2-DE. RESULTS： Rats in lowdose group didn＇ t suffer from remarkable renal impairment, while rats in high - dose group were successfully established as acute renal impairment models of cyclosporine. The protein of rat plasma after stepwise ammonium sulfate precipitation was satisfactorily separated so that protein dots between control group and high - dose group could be remarkably differentiated. CONCLUSION： The protocol can be applied to the search for protein biomarkers of cyclosporine renal toxicity.

关 键 词：环孢素 肾毒性 血浆 蛋白质标识物 双向电泳法 大鼠 

分 类 号：R965[医药卫生—药理学]