尿蛋白质组学在疾病标志物研究中的应用

尿液是重要的疾病标志物来源. 本文介绍了当前尿蛋白质组学的研究进展和尿液中疾病标志物研究的主要问题, 并对未来的发展进行了展望. 由于实际的临床问题通常是对症状相似的多种疾病进行鉴别诊断, 仅仅比较某一种疾病组和健康人对照组的尿蛋白质组差异不足以找到具有诊断能力的标志物. 另外, 尿蛋白质组在个体间和同一个体的不同生理条件下的变化也为疾病标志物的寻找带来了困难. 本文提出, 进行正常人群个体间和不同生理条件下尿蛋白质变化范围的研究可以为鉴定疾病标志物提供参考标准, 从而帮助研究者发现由疾病、而不是生理学差异引起的蛋白的变化. 比较蛋白在血浆和尿液中丰度的变化可以揭示肾脏的生理学功能和发现疾病标志物. 最后提出, 建立一个数据共享平台, 收集和整合已有的疾病标志物研发成果, 将大大推动尿蛋白质组研究的发展.     

激光显微切割/质谱联用技术在肾脏疾病诊断中的应用进展

激光显微切割(Laser microdissection,LMD)质谱(Mass spectrometry,MS)联用技术(LMD/MS)已成功应用于肾活检组织甲醛固定石蜡包埋切片的蛋白质组学研究,提高了某些肾脏病的诊断水平,显示出较好的临床应用前景。文中就LMD/MS蛋白质组学技术的原理、方法及该技术在肾淀粉样变性、膜增殖性肾小球肾炎等肾脏疾病的发病机制及诊断分型的应用进展进行综述。     

尿液蛋白质组在膀胱癌临床诊治中的应用

膀胱癌是一种常见的泌尿系统疾病,尿细胞学检查与膀胱镜检查是膀胱癌的主要临床诊断手段,但尿细胞学检查敏感性较差,膀胱镜检查为侵入性检查,易给病人带来强烈的不适感;且膀胱癌具有易复发的特点,大部分患者必须面临频繁的检查,临床亟需发展舒适、准确的检查手段.尿液存储是膀胱的主要生理作用,尿液可以直接接触肿瘤实体,肿瘤分泌的一些蛋白质分子极可能进入尿液中,并且患者尿液样本便于足量多次收集.同时,蛋白质组技术以及尿液蛋白质组研究的快速发展,为我们利用尿液研究膀胱癌提供了便利的途径.本文系统总结了尿液蛋白质组研究的主要技术手段,重点关注膀胱癌尿液蛋白质组研究趋势和应用方向,以期为利用尿液蛋白质组研究膀胱癌提供助力.     

尿液诊断膀胱癌的研究进展

摘要：膀胱癌是临床常见的发生在泌尿系统的恶性肿瘤,该病的发病率呈现逐年升高的趋势,其复发率也相对较高。早期诊断和定期随访是保证膀胱癌患者长期生存的关键。对于膀胱癌的诊断以及患者的随访通常凭借膀胱镜检查或尿脱落细胞学的测定。然而,前者的检查费用较为昂贵,且属于有创诊断;后者则具有检查敏感性相对较低的特点,还存在较大程度受病理科诊断医生主观因素影响的局限,目前还没有尿液生物标志物可以替代传统的诊断方法。膀胱肿瘤具有广泛的异质性,不同的疾病表型具有不同的分子差异。因此,引入尿液生物标志物来诊断疾病,评估疾病的侵袭性、进展的风险、复发的可能性和预后具有重要的临床价值。本文总结了目前尿液所含生物标志物诊断膀胱癌的研究现状,并对此领域的主要研究进展进行综述。     

蛋白质组学-引领后基因组时代

蛋白质组学是建立在高通量筛选技术的基础上发展的方法学,用于研究细胞功能网络模块中蛋白相互作用及在疾病或病变中蛋白和蛋白相互作用所发生的系统动态的差异变化;其研究技术奠基于双向凝胶电泳。及至世纪之交,随着质谱及蛋白质芯片的引进,蛋白质组学已广泛应用在生命科学上。其在医学上的应用,主要旨在发现疾病的特异性蛋白质分子或其蛋白质纹印,以揭示疾病的发生机制,也作为早期诊断、分子分型、疗效及预后判断的依据,并找出可能成为新药物设计的分子靶点,为疾病提供新的治疗方案。随着人类基因序列的完成,蛋白质组学热浪掀起了后基因组年代的序幕,人类将更深入地了解疾病和生命的本源。现就蛋白质组学10年来的发展历程、研究技术、在人类疾病中的应用及未来展望等作出精简的评述。     

慢性肾脏病尿液生物标志物研究进展:聚焦RNA转录标志物

慢性肾脏病(chronic kidney disease, CKD)已经越来越成为世界范围内的主要疾病负担之一,然而,目前仍缺乏早期诊断及动态监测其发生和发展的生物标志物。因此,在未来的精准医疗时代,寻找敏感、无创诊断标志物已经成为CKD诊疗所面临的重要挑战。本文将从尿液的沉渣、上清、细胞外囊泡三个方面,聚焦基因转录标志物(miRNA, mRNA),论述CKD尿液生物标志物的研究进展及尿液基因转录标志物研究所面临的挑战和未来可能的研究方向。     

蛋白质组学在妊娠期高血压疾病中的研究进展

蛋白质组学的兴起能快速筛选并鉴定疾病的特异性生物标志物,有助于研究妊娠期高血压疾病的病因、发生机制,通过特异性生物标志物进行早期诊断,从而改善母儿结局,降低母儿严重发病率和死亡率。本文综述近年来与妊娠期高血压疾病相关的蛋白质研究及其在妊娠期高血压疾病患者血液、脑脊液、尿液、羊水、滋养细胞中差异蛋白质谱的分析,为进一步研究提供思路。     

血浆蛋白质组学的研究进展

血浆蛋白质组学是研究血浆蛋白质的功能和变化的一门科学。血浆中蕴藏着生命机体的所有信息,因此只有彻底了解血浆中存在哪些蛋白质,才能知道如何利用血浆来预测人体对疾病的易感性并监控疾病的进程,以期达到对疾病进行早诊断早治疗。由于血浆蛋白质组动态范围大,给研究带来了很大的困难。尤其是高丰度蛋白质的存在影响了低丰度蛋白质的检测率。而低丰度蛋白质都是有意义的具有临床诊断价值的蛋白质。因此去除高丰度蛋白质的干扰成了血浆蛋白质组学研究的关键。近年来,血浆蛋白质组学相关研究技术也得到了长足进展,为深入研究血浆蛋白质做出了重要贡献。血浆蛋白质组学作为一种无创性的研究方法,值得我们去探讨。本文就血浆蛋白质组学研究进展情况做一简要综述。     

实验比格犬群发性泌尿系统结石的诊治

目的寻找某实验比格犬繁育基地群发性泌尿系统结石的病因及其有效的防治措施。方法采用临床诊断、病理诊断、结石成分分析、饲料成分分析、回顾性调查分析等手段以及综合的处理措施。结果经分析发现该群发性泌尿系统结石是由肉粉引起的尿酸盐结石,通过碱化尿液、降低血尿酸和去除病因等方法后,全群得到了有效的治疗。结论该肉粉是引起此次群发性泌尿系统结石的主要原因,由尿酸盐引起的、未发生器质性病变的尿路结石症可以治愈。     

尿外泌体在肾脏疾病中的研究进展

外泌体是体内多种细胞分泌的30-100nm的脂质双层膜结构的囊泡,内含蛋白质、核酸、脂质等成分。尿外泌体在生理状态下主要来源于面向管腔的各种细胞,尿外泌体内载的蛋白质、microRNA等参与泌尿系统微环境中细胞间的信息交流,与肾脏疾病发生、发展密切相关。检测尿外泌体特定蛋白或microRNA、mRNA的表达,可以反映特定的疾病,有利于肾脏疾病的诊断和鉴别诊断,甚至判断预后。本文就尿外泌体在肾脏疾病中的研究进展进行综述。     

Proteomic analysis of renal diseases: unraveling the pathophysiology and biomarker discovery

Current biomedical applications of proteomics have been conducted with four main objectives: to better understand the normal biology and physiology of cells, microorganisms, tissues and organs; to explore the pathogenic mechanisms and better understand the pathophysiology of medical diseases; to identify novel biomarkers for early disease detection, prediction and prognosis; and to define new therapeutic targets, drugs and vaccines. This review focuses predominantly on proteomic applications to unravel the pathophysiology and to define novel biomarkers for various renal diseases (i.e., glomerular diseases, tubulointerstitial diseases, renal vascular disorders and renal cancers). In addition, proteomic evaluations of renal transplantation and renal replacement therapy (for acute renal failure and end-stage renal disease) are summarized. Personal opinion, future perspectives and information resources for the field of renal and urinary proteomics are provided.     

Proteomic analysis of renal diseases: unraveling the pathophysiology and biomarker discovery

Current biomedical applications of proteomics have been conducted with four main objectives: to better understand the normal biology and physiology of cells, microorganisms, tissues and organs; to explore the pathogenic mechanisms and better understand the pathophysiology of medical diseases; to identify novel biomarkers for early disease detection, prediction and prognosis; and to define new therapeutic targets, drugs and vaccines. This review focuses predominantly on proteomic applications to unravel the pathophysiology and to define novel biomarkers for various renal diseases (i.e., glomerular diseases, tubulointerstitial diseases, renal vascular disorders and renal cancers). In addition, proteomic evaluations of renal transplantation and renal replacement therapy (for acute renal failure and end-stage renal disease) are summarized. Personal opinion, future perspectives and information resources for the field of renal and urinary proteomics are provided.     

Aldosterone as a renal growth factor

Aldosterone regulates blood pressure through its effects on the cardiovascular system and kidney. Aldosterone can also contribute to the development of hypertension that leads to chronic pathologies such as nephropathy and renal fibrosis. Aldosterone directly modulates renal cell proliferation and differentiation as part of normal kidney development. The stimulation of rapidly activated protein kinase cascades is one facet of how aldosterone regulates renal cell growth. These cascades may also contribute to myofibroblastic transformation and cell proliferation observed in pathological conditions of the kidney. Polycystic kidney disease is a genetic disorder that is accelerated by hypertension. EGFR-dependent proliferation of the renal epithelium is a factor in cyst development and trans-activation of EGFR is a key feature in initiating aldosterone-induced signalling cascades. Delineating the components of aldosterone-induced signalling cascades may identify novel therapeutic targets for proliferative diseases of the kidney.     

In search of adult renal stem cells

The therapeutic potential of adult stem cells in the treatment of chronic degenerative diseases has becoming increasingly evident over the last few years. Significant attention is currently being paid to the development of novel treatments for acute and chronic kidney diseases too. To date, promising sources of stem cells for renal therapies include adult bone marrow stem cells and the kidney precursors present in the early embryo. Both cells have clearly demonstrated their ability to differentiate into the kidney's specialized structures. Adult renal stem cells have yet to be identified, but the papilla is where the stem cell niche is probably located. Now we need to isolate and characterize the fraction of papillary cells that constitute the putative renal stem cells. Our growing understanding of the cellular and molecular mechanisms behind kidney regeneration and repair processes - together with a knowledge of the embryonic origin of renal cells - should induce us, however, to bear in mind that in the kidney, as in other mesenchymal tissues, the need for a real stem cell compartment might be less important than the phenotypic flexibility of tubular cells. Thus, by displaying their plasticity during kidney maintenance and repair, terminally differentiated cells may well function as multipotent stem cells despite being at a later stage of maturation than adult stem cells. One of the major tasks of Regenerative Medicine will be to disclose the molecular mechanisms underlying renal tubular plasticity and to exploit its biological and therapeutic potential.     

Lysophosphatidic acid and renal fibrosis

The development of fibrosis involves a multitude of events and molecules. Until now the majority of these molecules were found to be proteins or peptides. But recent data show significant involvement of the phospholipid lysophosphatidic acid (LPA) in the development of pulmonary, liver and renal fibrosis. The latest data on the role of LPA and the G-protein-coupled LPA1 receptor in the development of renal fibrosis will be discussed. LPA1-receptor activation was found to be associated with increased vascular leakage and increased fibroblast recruitment in pulmonary fibrosis. Furthermore, in renal fibrosis LPA1-receptor activation stimulates macrophage recruitment and connective tissue growth factor expression. The observations make this receptor an interesting alternative and new therapeutic target in fibrotic diseases.     

Therapeutic application of extracellular vesicles for various kidney diseases: a brief review

Extracellular vesicles (EVs) released from different types of kidney cells under physiologic conditions contribute to homeostasis maintenance, immune-modulation, and cell-to-cell communications. EVs can also negatively affect the progression of renal diseases through their pro-inflammatory, pro-fibrotic, and tumori-genic potential. Inhibiting EVs by blocking their production, release, and uptake has been suggested as a potential therapeutic mechanism based on the significant implication of exosomes in various renal diseases. On the other hand, stem cell-derived EVs can ameliorate tissue injury and mediate tissue repair by ameliorating apoptosis, inflammation, and fibrosis while promoting angiogenesis and tubular cell proliferation. Recent advancement in biomedical engineering technique has made it feasible to modulate the composition of exosomes with diverse biologic functions, making EV one of the most popular drug delivery tools. The objective of this review was to provide updates of recent clinical and experimental findings on the therapeutic potential of EVs in renal diseases and discuss the clinical applicability of EVs in various renal diseases.     

The emerging role of cellular senescence in renal diseases

Cellular senescence represents the state of irreversible cell cycle arrest during cell division. Cellular senescence not only plays a role in diverse biological events such as embryogenesis, tissue regeneration and repair, ageing and tumour occurrence prevention, but it is also involved in many cardiovascular, renal and liver diseases through the senescence‐associated secretory phenotype (SASP). This review summarizes the molecular mechanisms underlying cellular senescence and its possible effects on a variety of renal diseases. We will also discuss the therapeutic approaches based on the regulation of senescent and SASP blockade, which is considered as a promising strategy for the management of renal diseases.     

Detection of renal cell carcinoma-associated markers via proteome- and other 'ome'-based analyses.

Proteome analysis has rapidly developed in the post-genome era and is now widely accepted as the complementary technology for genetic profiling. It has been shown to be a powerful tool for studying human diseases and for identifying novel prognostic, diagnostic and therapeutic markers. This review focuses on the identification of new biomarkers and therapeutic targets for renal cell carcinoma using different 'ome'-based technologies.     

足细胞发育异常及相关肾脏疾病研究进展

足细胞是肾小球滤过屏障的重要组成部分,其数量减少或功能障碍将导致肾小球滤过功能损伤和相关肾脏疾病的发生。足细胞为不可再生性细胞,其数量和功能在一定程度上取决于其正常发育。已发表的文献和本实验室的研究工作表明,遗传或不良宫内环境等原因所致的足细胞发育不良,可能导致成年后肾小球滤过功能障碍,并成为某些胎源性肾脏疾病发生或易感的病因之一,而表观遗传学机制可能参与介导足细胞发育过程中某些关键基因的表达异常。本文对足细胞结构功能和正常发育、足细胞发育异常的病因和机制、以及足细胞发育异常所致的肾脏疾病等几方面进行综述,以期对发育源性足细胞相关肾脏疾病的诊断与治疗提供借鉴与参考。     

Renal fibrosis: insight from proteomics in animal models and human disease

Chronic kidney disease (CKD) is the end-point of a number of renal and systemic diseases. The high incidence and financial burden of CKD makes it imperative to diagnose CKD at early stages when therapeutic interventions are far more effective. A key component of CKD is the development of renal fibrosis. Renal fibrosis is a complex process, associated with many cell types and pathways, resulting in structural and functional alterations. Identification of specific biomarkers of renal fibrosis may thus not only help us to understand the pathophysiological mechanisms involved in this process, but also improve diagnosis in the clinic. In this review, the existing literature on proteomic approaches to study renal fibrosis is presented and evaluated. The importance of using animal models along with patient material is discussed and future directions, considered key to this field, are proposed.