COMPARATIVE PROTEOMIC INVESTIGATION OF ARDS AND SYSTEMIC INFLAMMATORY INJURY

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。急性呼吸窘迫综合征（acute respiratory distress syndrome， ARDS）是重症监护医学中最重要的发病和死亡原因之一。它可以是许多疾病的后遗症，如败血症、胃内容物误吸、肺炎或重大创伤。该综合征的特点是炎症反应导致肺泡-毛细血管屏障破裂，导致血液中的液体和蛋白质流入肺泡空间。炎症反应的确切机制尚不完全清楚。为了提高对这种疾病状态的理解和评估可能的治疗方案，已经进行了大量的临床和实验试验。方法：本研究将ARDS患者的肺水肿液样本与心源性肺水肿患者的对照样本进行比较。通过层析、过滤和电泳分离蛋白质后，将使用质谱（MALDI-TOF-TOF， ESI）分析两组蛋白质组和蛋白质翻译后修饰，以确定疾病的蛋白质标记物。最后，通过免疫分析进一步评估浓度变化。结果：通过柱层析和过滤，肺水肿液样品清除了碎片，1D SDS-PAGE可分离蛋白质。目前正在评价MALDI-TOF-TOF的结果。下周将进行高压液相色谱和ESI分析。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The acute respiratory distress syndrome (ARDS) is one of the most important causes for morbidity and mortality in intensive care medicine. It can be the sequel of numerous diseases like sepsis, aspiration of gastric contents, pneumonia or major trauma. The syndrome is characterized by an inflammatory reaction that leads to a breakdown of the alveolar-capillary barrier, resulting in an influx of fluid and proteins from the blood into the alveolar space. The exact mechanism of the inflammatory reaction is still incompletely understood. Numerous clinical and experimental trials have been made in order to improve the understanding and evaluate possible treatment options of this disease state. METHODS: The proposed study focuses on the evaluation of pulmonary edema fluid samples of patients with ARDS compared to control samples from patients with cardiogenic pulmonary edema. After protein separation by chromatography, filtration and electrophoresis, mass spectrometry (MALDI-TOF-TOF, ESI) will be used to analyze the proteome and posttranslational modifications of proteins in both groups in order to identify protein markers of disease. Eventually, concentration changes shall be further evaluated by immunoassays. RESULTS: Using column chromatography and filtration, the pulmonary edema fluid samples were cleaned from debris and proteins could be separated by 1D SDS-PAGE. The results of MALDI-TOF-TOF are currently being evaluated. Analysis by High Pressure Liquid Chromatography and ESI is scheduled for next week.