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。慢性肾脏病是医学界高度重视的疾病，作为慢性肾脏病患者的一部分，将来会失去肾功能，需要透析治疗或肾移植。病理检查可将本病分为多种疾病，但其发病机制和病理生理机制尚不清楚，这给治疗CKD带来了困难。本研究旨在通过对CKD患者肾活检获得的肾小球组织标本进行生物信息学分析，了解其发病机制和病理生理机制。首先，利用质谱仪(MS)和抗体对正常人肾小球蛋白质组进行蛋白质组学分析，以了解正常人肾小球蛋白质组。用筛选法从肾癌患者的肾脏中分离出肾小球。在肾小球中鉴定了3000多种基因衍生蛋白，结果通过人类肾小球蛋白质组数据库(http：//www.香港大学。第二，将MS蛋白质组学应用于冰冻或福尔马林固定的石蜡包埋(FFPE)肾活检标本。通过激光显微切割法从每个肾活检标本中收集了50个肾小球切片，并计算了蛋白质或多肽的数量。

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