蛋白质组学在消化系疾病中医药研究中的应用

自然界各种生命物质都是由蛋白质组成的,各种蛋白质按不同的组成、结构和修饰,组成各种细胞和生命物质.蛋白质组学通过从整体角度分析细胞内动态变化的蛋白质组成、表达水平、修饰状况、蛋白质之间以及蛋白质与其他生物分子之间的相互作用,从而了解蛋白质的功能及其在生命过程中的作用.它包括双向凝胶电泳、生物质谱鉴定、生物信息学.当人体中某部分发生生理、病理变化时,该机体组织的蛋白质组发生变化,或代谢物的蛋白质组发生变化,就可以通过蛋白质组学技术找到这些新产生的蛋白质(称为"上调"或"表达增高)或减少甚至消失的蛋白质(称为"下调"或"表达降低"),这些蛋白质叫做"差异蛋白",是用于研究生理、病理变化诊断的潜在"靶点".     

蛋白质组学在糖尿病肾病研究中的应用现状

蛋白质组学是"后基因组时期"生物医学研究的重点内容之一。它是一门对某一细胞或机体在特定的生理或病理状态下表达的所有蛋白质的特征、数量和功能进行系统性研究的科学。本文简要介绍了蛋白质组学的研究内容及技术平台,并对目前蛋白质组学技术在糖尿病肾病研究中的应用现状作一综述。     

蛋白质组学技术在恶性肿瘤生物标志物探索中的应用

蛋白质组由澳大利亚学者Wilkins和Williams最先提出，包括细胞内基因组编码的全套蛋白质。蛋白质组学研究从细胞水平及整体水平研究蛋白构成及其变化规律，对不同生理或病理条件下蛋白表达的差异进行比较；可对特定时间和环境下细胞或组织蛋白质组进行高通量研究，为发现特定生理或病理条件下蛋白质组发生的变化提供新的技术平台，有利于发现用于早期诊断的新的恶性肿瘤生物标志物，提高恶性肿瘤的治疗水平。现将近年来蛋白质组学技术在恶性肿瘤生物标志物探索中的应用进展综述如下。     

男性高血压前期与正常血压及原发性高血压患者血浆比较蛋白质组学研究

目的 探讨正常血压、高血压前期及高血压患者血浆蛋白质组学差异.方法 筛选正常血压者,高血压前期及高血压男性患者各26例,分离血浆,去除高丰度蛋白质,制备蛋白质样品,通过使用二维电泳结合基质辅助激光解析电离飞行时间质谱技术进行各组血浆蛋白质组的分析和鉴定,并通过Western blot进一步进行验证.结果 在三组不同血压的患者血浆中存在着22个表达差异的蛋白质点,这些点与18种蛋白质在相关.主要涉及炎症与免疫、脂质代谢、转运蛋白质、凝血和纤溶、细胞增殖和凋亡以及抗氧化蛋白质.结论 高血压前期患者血浆蛋白质组与正常血压及高血压患者之间存在明显的差别,高血压组也与正常血压组不同.高血压前期可能是高血压早期的一种表现.     

心血管疾病线粒体蛋白质组学研究

线粒体是真核细胞重要的细胞器,线粒体功能障碍导致很多疾病包括心血管疾病。随着蛋白质组学技术的不断进步,从整体上研究不同病理状态下线粒体内蛋白质的含量变化渐受关注。本文综述线粒体蛋白质组学的主要技术方法进展及其在心血管疾病研究中的应用。     

心血管疾病线粒体蛋白质组学研究

线粒体是真核细胞重要的细胞器，线粒体功能障碍导致很多疾病包括心血管疾病。随着蛋白质组学技术的不断进步，从整体上研究不同病理状态下线粒体内蛋白质的含量变化渐受关注。本文综述线粒体蛋白质组学的主要技术方法进展及其在心血管疾病研究中的应用。     

蛋白质组学及其在肝癌中的应用

蛋白质组学是以蛋白质组为研究对象的一门新兴学科,其研究技术主要包括双向凝胶电泳、质谱和生物信息学。蛋白质组学在肝癌中有广泛应用,包括寻找肝癌早期诊断的生物标记、判断肝癌是否转移及术后复发等。     

蛋白质组学在胰腺疾病研究中的应用

蛋白质组学是研究蛋白质的表达、翻译后修饰、在细胞内定位以及蛋白质与蛋白质之间的相互作用的科学。传统的研究手段主要采用双向凝胶电泳和质谱技术,前者用于蛋白质的分离,后者用于蛋白质的鉴定。目前新兴的研究技术主要有相差凝胶电泳技术同位素亲和标签技术。蛋白质组学在胰腺疾病的研究主要是通过正常个体与患病个体的血清、胰液或组织等进行蛋白质比较,发现差异蛋白质,从而对急、慢性胰腺炎和胰腺癌等胰腺疾病的发病机制、早期诊断和有效治疗提供理论依据和新思路。     

肿瘤蛋白质组学与消化系肿瘤生物标记

肿瘤蛋白质组学是通过蛋白质组技术研究肿瘤细胞中蛋白质及其相互作用的新的学科。将蛋白质组学技术用于研究肿瘤的发病机制、寻找新的肿瘤诊断生物标记以及采用组织蛋白质组图谱进行早期诊断等早已引起了人们的广泛关注。肿瘤蛋白质组学可能会引起肿瘤临床实践的重大革命。本文对近年发现的肿瘤生物标记进行了回顾,分析了肿瘤蛋白质组学在肿瘤生物标记发展中的前景。     

微粒蛋白质组学研究进展

1994年澳大利亚学者Wilkins等最早提出蛋白质组的概念,它是指由一个基因组或者一个细胞、组织在某一特定时刻表达的包括所有亚型和修饰的蛋白质的总和。蛋白质组学是从整体上分析细胞内的蛋白质种类、数量、修饰状态,了解蛋白质的功能与相互作用。与基因组相比,蛋白质是生物体执行其生物功能的基本单位,因此蛋白质组对生     

刚地弓形虫蛋白质组研究进展

随着基因组学的发展和完善,在基因组学的基础上,蛋白质组学作为一门新兴的前沿科学已经显示出了其巨大的发展前景.将蛋白质组学技术应用于弓形虫研究,在蛋白质水平上全面认识弓形虫的生理、病理等过程是目前弓形虫研究领域的热点之一.该文就弓形虫蛋白质组的研究进展进行综述,以期从蛋白质组学研究的角度为弓形虫研究提供新的思路.     

Clinical proteomics: current status, challenges, and future perspectives

This account will give an overview and evaluation of the current advances in mass spectrometry (MS)-based proteomics platforms and technology. A general review of some background information concerning the application of these methods in the characterization of molecular sizes and related protein expression profiles associated with different types of cells under varied experimental conditions will be presented. It is intended to provide a concise and succinct overview to those clinical researchers first exposed to this foremost powerful methodology in modern life sciences of postgenomic era. Proteomic characterization using highly sophisticated and expensive instrumentation of MS has been used to characterize biological samples of complex protein mixtures with vastly different protein structure and composition. These systems are then used to highlight the versatility and potential of the MS-based proteomic strategies for facilitating protein expression analysis of various disease-related organisms or tissues of interest. Major MS-based strategies reviewed herein include (1) matrix-assisted laser desorption ionization-MS and electron-spray ionization proteomics; (2) one-dimensional or two-dimensional gel-based proteomics; (3) gel-free shotgun proteomics in conjunction with liquid chromatography/tandem MS; (4) Multiple reaction monitoring coupled tandem MS quantitative proteomics and; (5) Phosphoproteomics based on immobilized metal affinity chromatography and liquid chromatography-MS/MS.     

Application of the yeast two-hybrid system in molecular gerontology

Most – if not all – proteins are bound to interact with other proteins to exert their function, and thus the identification of the interaction partners of a protein is vital in proteomics. The yeast two-hybrid system is a popular and effective tool for studyingprotein–protein interactions. Although the advantages of the system are manifold, it also has certain drawbacks and limitations. The two-hybrid system has been shown to be extremely useful for placing a protein of unknown function within a functional context, thereby providing information about a putative role of the uncharacterised protein. This concept has also been successfully applied in molecular gerontology. This revised version was published online in July 2006 with corrections to the Cover Date.     

Precision proteomics: the case for high resolution and high mass accuracy

Proteomics has progressed radically in the last 5 years and is now on par with most genomic technologies in throughput and comprehensiveness. Analyzing peptide mixtures by liquid chromatography coupled to high-resolution mass spectrometry (LC-MS) has emerged as the main technology for in-depth proteome analysis whereas two-dimensional gel electrophoresis, low-resolution MALDI, and protein arrays are playing niche roles. MS-based proteomics is rapidly becoming quantitative through both label-free and stable isotope labeling technologies. The latest generation of mass spectrometers combines extremely high resolving power, mass accuracy, and very high sequencing speed in routine proteomic applications. Peptide fragmentation is mostly performed in low-resolution but very sensitive and fast linear ion traps. However, alternative fragmentation methods and high-resolution fragment analysis are becoming much more practical. Recent advances in computational proteomics are removing the data analysis bottleneck. Thus, in a few specialized laboratories, “precision proteomics” can now identify and quantify almost all fragmented peptide peaks. Huge challenges and opportunities remain in technology development for proteomics; thus, this is not “the beginning of the end” but surely “the end of the beginning.”     

尿液蛋白质组学与糖尿病肾病

蛋白质组学是系统研究细胞或组织全套蛋白质生物学信息的科学,其中尿液蛋白质组学以其独特的优点广泛应用于基础和临床研究领域.糖尿病肾病是糖尿病最常见且危害巨大的并发症之一,许多学者运用尿液蛋白质组学分析方法对糖尿病肾病进行研究,发现了一些与糖尿病肾病相关的生物学标志及可能的发生机制,展示了尿液蛋白质组学技术在糖尿病肾病早期诊断、动态监测病情及发现新的治疗靶点方面的前景.     

弥漫性间质性肺疾病蛋白组学研究

蛋白质组学是指对蛋白质组整体水平进行研究.与传统研究单一蛋白质变化的方法 发相比,能获得整体蛋白质水平表达差异.弥漫性间质性肺疾病发病机制复杂,涉及广泛的细胞因子网络,以往大量研究集中于单个细胞因子水平,蛋白质组学技术以其高新系统量及整体水平比较差异优势成为新的研究方向.本文就蛋白质组学技术在肺间质疾病的研究应用进行综述.     

Differential proteomic analysis of human colorectal carcinoma cell lines metastasis-associated proteins

Purpose Metastasis is a common phenomenon and the major lethal cause of colorectal carcinoma (CRC). To better comprehend the mechanism underlying CRC metastasis and to search for potential markers for predicting CRC metastasis, two CRC cell lines with different metastatic potentials, SW480 and SW620, were investigated by phenotypic analyses and proteomics technologies. Methods The surgical orthotopic implantation (SOI) technique was originally used to develop a reproducible colorectal cancer model in nude mice with stable tumor growth and metastasizing course. Furthermore, differential proteome analysis of two CRC cell lines was conducted using two-dimensional (2-D) gel electrophoresis combined with matrix-assisted laser desorption/time of flight (MALDI-TOF) mass spectrometry. Results Among the differential spots, 12 protein spots (11 proteins) were further identified using in-gel tryptic digestion and peptide mass fingerprinting (PMF). Interestingly, most of these proteins we identified have been reported to be associated with distinct aspect of tumor metastasis to some extent. Our immunohistochemistry assays of colorectal cancer revealed that heat shock protein (HSP) 27 overexpression relates to metastatic behavior of CRC cell. Conclusions The protein profile between two colorectal cell lines with different potential metastasis displayed obvious differences. The results imply that various different proteins may lead to CRC metastasis together. HSP27 overexpression played an important role in metastasis and progression of CRC.     

类风湿关节炎患者外周血单个核细胞蛋白质组学研究

目的 运用蛋白质组学的方法,比较正常人及早期活动性类风湿关节炎(RA)患者外周血单个核细胞(PBMCs)蛋白质的差异表达,寻找RA疾病相关致病蛋白质.方法 选取9例早期活动期RA患者以及9名健康成年志愿者,用淋巴细胞分离液分离PBMCs,抽提PBMCs中的蛋白,采用同相pH梯度(IPG)双向凝胶电泳(2-DE)分离正常人及RA患者PBMCs总蛋白质.凝胶经考马斯亮蓝染色显色后,PDQuest图像分析软件进行比较分析、识别差异表达的蛋白质,对差异蛋白质点用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)进行鉴定,运用反转录-聚合酶链反应(RT-PCR)方法验证部分差异蛋白质.结果 获得RA患者及正常人PBMCs蛋白质双向凝胶电泳图谱,平均蛋白质点数分别为556和579,匹配率分别为89.4%和 88.5%,通过比较分析,差异表达蛋白质点数为23,选取18个点进行质谱鉴定,成功鉴定14个蛋白质,其中a-肌动蛋白、纤维蛋白素原a-链、载脂蛋白A-I(ApoA-I)等9个蛋白质点在RA中表达上调,硫氧还蛋白-2、谷胱甘肽S-转移酶等6个蛋白质点在RA中表达下调,这些差异蛋白质的功能涉及物质代谢、抗氧化、信号传导、能量产生及细胞骨架.并用RT-PCR方法验证差异蛋白质ApoA-I.其结果与上述蛋白质差异表达结果相符.结论 在RA患者PBMCs中存在着差异表达蛋白质,这些差异蛋白质可能是RA发病的内在因素.其RT-PCR结果与蛋白质差异表达相符,证明蛋白质组研究的可靠性.     

肺结核患者外周血单个核细胞蛋白质差异的表达

目的 应用比较蛋白质组学方法筛选出肺结核患者发病免疫相关蛋白质,为阐明结核病的发病的免疫机制和诊断治疗提供理论依据.方法 收集10例肺结核患者和10名健康志愿者外周血单个核细胞(PBMC),采用双向凝胶电泳分离细胞总蛋白,运用Image Master 2D Elite 5.0图像分析软件识别差异表达的蛋白质点,应用基质辅助激光解析电离飞行时间质谱获取肽质量指纹图谱,检索数据库鉴定差异表达的蛋白质点,并对差异表达的蛋白质用Western blot进行验证.两组间比较采用秩和检验.结果 建立了重复性良好的肺结核患者和健康志愿者PBMC的双向凝胶电泳图谱,找出14个表达量有明显差异的蛋白质点并鉴定出共12种蛋白质,其中冠蛋白1A等10种蛋白在肺结核组中表达上调,普列克底物蛋白(PLEK)和Ras基因抑制蛋白1(RSU1)在肺结核组中表达下调;Western blot法验证PLEK在肺结核组表达下调,与双向凝胶电泳结果一致.结论 对肺结核患者PBMC的比较蛋白质组学研究鉴定出冠蛋白1A、PLEK等12种差异蛋白质,其中PLEK和冠蛋白1A可能通过对单核巨噬细胞吞噬作用的调节在肺结核发病中起着重要作用.     

Proteomics in Inflammatory Bowel Disease: Approach Using Animal Models

Recently, proteomics studies have provided important information on the role of proteins in health and disease. In the domain of inflammatory bowel disease, proteomics has shed important light on the pathogenesis and pathophysiology of inflammation and has contributed to the discovery of some putative clinical biomarkers of disease activity. By being able to obtain a large number of specimens from multiple sites and control for confounding environmental, genetic, and metabolic factors, proteomics studies using animal models of colitis offered an alternative approach to human studies. Our aim is to review the information and lessons acquired so far from the use of proteomics in animal models of colitis. These studies helped understand the importance of different proteins at different stages of the disease and unraveled the different pathways that are activated or inhibited during the inflammatory process. Expressed proteins related to inflammation, cellular structure, endoplasmic reticulum stress, and energy depletion advanced the knowledge about the reaction of intestinal cells to inflammation and repair. The role of mesenteric lymphocytes, exosomes, and the intestinal mucosal barrier was emphasized in the inflammatory process. In addition, studies in animal models revealed mechanisms of the beneficial effects of some therapeutic interventions and foods or food components on intestinal inflammation by monitoring changes in protein expression and paved the way for some new possible inflammatory pathways to target in the future. Advances in proteomics technology will further clarify the interaction between intestinal microbiota and IBD pathogenesis and investigate the gene-environmental axis of IBD etiology.