Analysis of pathogenesis and pathophysiology of chronic kidney disease glomerulus by microproteomics

慢性肾脏病（CKD）的主要病理损害是肾小球。由于部分慢性肾脏病患者将来会丧失肾功能，需要进行透析治疗或肾移植，因此慢性肾脏病受到医学界的高度重视。本研究旨在通过生物信息学方法分析CKD患者肾活检获得的肾小球组织标本，了解CKD的发病机制和病理生理。为了了解正常人肾小球蛋白质组，首先利用质谱仪和抗体对正常人肾小球蛋白质组进行了全面的蛋白质组学研究。从肾癌患者的肾脏中通过筛分法分离肾小球。在肾小球中鉴定了超过3000种基因衍生的蛋白质，并且结果通过作为人类肾小球蛋白质组数据库的网站（http：//www. hkupp。其次，将MS蛋白质组学应用于冷冻或福尔马林固定石蜡包埋（FFPE）肾活检样品。通过激光显微切割方法，从每个肾活检标本中实际上可收集50个肾小球切片，并测定蛋白质或肽的量。

Primary pathological lesion of chronic kidney disease(CKD) is the glomerulus. The CKD is paid a serious attention in medicine as a part of CKD patients will lose their kidney function in the future and need dialysis treatment or kidney transplantation. This disease is classified into several diseases by pathological examination, however, the pathogenesis and pathophysiology are almost unknown, which make it difficult to develop curative treatments for CKD.The current study was aimed to analyze glomerular tissue samples obtained by kidney biopsy from CKD patients to understand the pathogenesis and pathophysiology by bioinformatics. At first, comprehensive proteome of normal human glomerulus was examined by proteomics using mass spectrometers(MS) and antibodies to know normal human glomerulus proteome. The glomeruli were isolated by the sieving method from kidneys removed from patients with renal carcinomas. More than 3000-gene-derived proteins were identified in the glomerulus and the results are opened for public through a website as a Human Glomerulus Proteome Database(http ://www. hkupp. org).Secondary, MS proteomics was applied to frozen or formalin-fixed paraffin-embedded(FFPE) kidney biopsy samples. Fifty glomerular sections were practically collectable by a laser micro dissection method from each kidney biopsy specimen and the amount of proteins or peptides