阿霉素肾病大鼠尿蛋白质组质谱的检测及中药肾康灵的干预作用  

A Urinary Proteomics Mass Spectromentry Detection and Chinese Intervention of Shenkangling

作　　者：艾斯[1] 郑健[1] 林青[1] 吴华嵩[2] 陈蓉艳[1] 陈乃清[1] 陈文列[3] 张洪生[1] 

机构地区：[1]福建中医学院附属人民医院,福州350004 [2]福建省中医药研究院,福州350003 [3]福建中医学院,省高校中西医结合基础重点实验室,福州350108

出　　处：《中国中西医结合肾病杂志》2010年第5期398-403,I0001,共7页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：福建省自然科学基金资助项目(No.2007J0307;2008J0093)

摘　　要：目的:应用尿蛋白质组学筛选阿霉素肾病(AN)大鼠尿液中差异表达的蛋白质,并检测中药肾康灵的干预作用,旨在蛋白质分子水平探索频复发性肾病(FRNS)的诊断标记物及药物治疗的作用靶点。方法:用阿霉素(ADR)5.5mg/kg诱导大鼠复制的阿霉素肾病(AN)模型,即类似于人类微小病变型(MCD)肾病模型。实验共分为5组,分别为正常组(A组)、AN病模组(B组)、泼尼松治疗组(C组)、泼尼松+肾康灵(低浓度)治疗组(D组)、泼尼松+肾康灵(高浓度)治疗组(E组)。利用表面增强激光解析-电离飞行时间质谱(SELDI-TOF-MS)技术联合CM10蛋白质芯片技术检测各组大鼠尿蛋白质组质谱的表达,采用PBSⅡ-C型蛋白质芯片阅读机自动收集数据,使用Ciphergen Proteinchip Software3.2.1和Biomarker Wizard分析软件筛选出各组大鼠尿液差异表达的蛋白质。并观察各组治疗前后尿蛋白、血清白蛋白(Alb)、胆固醇(TC)、三酰甘油(TG)、尿素氮(BUN)、血清肌酐(Scr)及肾脏病理形态学等变化。结果:在质荷比值(M/Z)2000～50000范围内(P<0.05),芯片共捕获到近40种差异蛋白质,质谱仪筛选出B组与A组比较的38种差异蛋白,其中3种差异蛋白从B组→C组→D、E组→A组呈递减趋势。这3种差异蛋白峰的M/Z值分别为9321.0、9393.1、9477.5。结论:SELDI-TOF-MS能有效分析阿霉素肾病大鼠尿液蛋白质质谱,中药肾康灵干预治疗后蛋白质质谱表达差异有统计学意义,且中药肾康灵联合泼尼松治疗AN大鼠的疗效优于单用泼尼松治疗。Objective：To screening the urinary proteomic patterns of rats with adriamycin-induced nephropathy by urinary proteomics for discussion of the biomarker of Frequent Relapse Nephrotic Syndrome and target point of medication.Methods：We copied adriamycin-induced nephropathy （AN） rats model with adriamycin （ADR） 5.5 mg/kg.All rats were divided into 5 groups：the normal group（group A）,the adriamycin-induced nephropathy group （group B）,the prednisone group （group C）,the prednisone and the Shenkangling（light concentration） group （group D）,the prednisone and Shenkangling （high concentration） group（group E）.Surface enhanced laser desorption/ionization-time of flight-mass spectrometry （SELDI-TOF-MS） CM10 protein clips were used to detect the urinary proteomic patterns of every rat.PBS Ⅱ-C protein clips reading machine was applied to collect data automatically.Proteins with differential expression between each group were analyzed by Ciphergen Proteinchip Software 3.2.1 and Biomarker Wizard software. And changes were observed on urinary protein,serum albumin （Alb）,cholesterol （TC）,triglyceride （TG）,blood urea nitrogen （BUN）,serum creatinine （Scr） and renal pathology in each group before and after treatment.Results：About 40 proteins with differential expressions were detected in the mass charge ratio range of 2 000～50 000 （P〈0.05）,there were 38 proteins between group B and group A,3 proteins decreased progressively from the B group,to the C group,the D group to the E group and to the A group.M/Z were 9 321.0、9 393.1、9 477.5.Conclusion：The urinary proteomic patterns of rats with adriamycin-induced nephropathy can be analyzed effectively by SELDI-TOF-MS.The difference of proteomics was significant before and after intervention of decoction Shenkangling.Traditional Chinese medicine Shenkangling combined prednisone was more effectively than prednisone alone in the treatment of AN rats.

关 键 词：SELDI-TOF-MS技术 CM10芯片 肾康灵 阿霉素肾病 尿蛋白质组学 

分 类 号：R692.9[医药卫生—泌尿科学] R285.5[医药卫生—外科学]