蛋白质组学及其在肾脏病研究中的应用幻灯片.ppt

This study was aimed at the search of urinary biomarkers which might help to predict the clinical response of IgA nephropathy (IgAN) patients to angiotensin converting enzyme inhibitors (ACEi). First, we studied the urinary proteome of 18 IgAN patients (toward 20 healthy controls) who had been chronically treated with ACEi by using 2-D PAGE coupled to nano-HPLC-ESI-MS/MS analysis. We identified 3 proteins, kininogen（激肽原） (p = 0.02), inter-alpha-trypsin-inhibitor heavy chain 4 (35kDa fragment) (p = 0.02) and transthyretin（甲状腺素转运蛋白）(p0.0001), whose urinary excretion was different in IgAN patients responders when compared to those who had not responded to ACEi. A reduction of daily proteinuria 50% and a stable renal function over time were used to classify patients as responders. Then, we adopted immunoblotting to confirm the predictive power of one of the above proteins, kininogen, in 20 patients with biopsy-proven IgAN, before starting any therapy. Thus, we confirmed that very low levels of kininogen urine excretion were indeed predictive of an inadequate or absent clinical response to ACEi therapy of IgAN patients, after 6-month follow-up. Concluding, the analysis of urine proteome of IgAN patients generated a set of proteins which distinguished subjects responsive to ACEi from those unresponsive to the inhibition of renin-angiotensin system (RAS). 黄艳军等对儿童微小病变型激素耐药与激素敏感型肾病综合征的尿中差异蛋白质表达进行了研究。 12个相关蛋白的鉴定将对激素耐药治疗靶位和耐药型肾病综合征诊断的分子标志物发现可能有所裨益。 -黄艳军，黄松明，张爱华，等.微小病变型肾病综合征激素耐药和敏感的比较尿蛋白质组学研究.南方医科大学学报, 2007；27(1 0) （四）中医药领域中的应用 证候学研究（整体性、复杂性、网络性） 中药及复方药理学研究(多组分，多环节，多靶点) 例：黄黎明,梁恒,邢建宇,等.山茱萸提取物对慢性肾炎小鼠的药理作用及蛋白质组学效应的实验研究.中国药理学通报, 2004 Oct；20(10)：1 l44～50. * 这就是将2DE和质谱相结合进行比较蛋白质组分析的实验方案。（方案完后），下面，我将汇报用以上介绍的方法进行脾淋巴细胞及乳腺癌比较蛋白质组学的实验结果。 2．噬菌体表面显示技术 ( phage display ) 三个基本因素： 在噬菌体pⅢ和pⅧ衣壳蛋白N端插入外源待筛基因编码的蛋白，形成的融合蛋白表达在噬菌体颗粒的表面；同时其保持的外源蛋白天然构象能被相应的抗体或受体识别 利用固相支持物上的抗体或受体，采用亲和筛选法（panning），从噬菌体文库中筛选出能结合筛选分子的目的噬菌体。 外源多肽或蛋白质表达在噬菌体的表面，其编码基因可通过分泌型噬菌体的单链DNA测序推导出来。 主 要 应 用 筛选与