Magi Barbara, Bini Luca, Perari Maria Grazia, Fossi Antonella, Sanchez Jean-Charles, Hochstrasser Denis, Paesano Salvo, Raggiaschi Roberto, Santucci Annalisa, Pallini Vitaliano, Rottoli Paola
Department of Molecular Biology, University of Siena, Italy.
Electrophoresis. 2002 Sep;23(19):3434-44. doi: 10.1002/1522-2683(200210)23:19<3434::AID-ELPS3434>3.0.CO;2-R.
We used two-dimensional (2-D) electrophoresis to analyze the protein composition of fluid recovered by bronchoalveolar lavage (BALF) from patients with sarcoidosis and idiopathic pulmonary fibrosis, two forms of interstitial lung disease with different cellular composition and cytokine profile in BALF. They are also characterized by different pathogenesis and clinical evolution, idiopathic pulmonary fibrosis being less favorable than sarcoidosis due to rapidly progressive pulmonary fibrosis. Thirty-eight proteins or protein fragments, never previously assigned in BALF samples, were identified by various methods including mass fingerprinting of tryptic digests. Comparison of the BALF protein maps of the two groups of patients showed 32 spots with statistically significant disease-related variations in relative abundance. In sarcoidosis we found an increase in the amount of several plasma proteins, while in idiopathic pulmonary fibrosis we observed a statistically significant increase in low-molecular-weight proteins, many of which are involved in inflammatory processes (such as MIF and calgranulin) or antioxidant response (such as antioxidant peroxysomal enzyme and thioredoxin peroxidase 2). 2-D electrophoresis allowed us to identify new BALF proteins and to characterize protein composition in patients with sarcoidosis and idiophatic pulmonary fibrosis. Comparison of the gels of the two diseases showed that they differ in BALF protein profiles as they do in type of immune response.
我们采用二维（2-D）电泳分析结节病和特发性肺纤维化患者支气管肺泡灌洗回收液（BALF）的蛋白质组成，这两种间质性肺疾病在BALF中的细胞组成和细胞因子谱不同。它们还具有不同的发病机制和临床病程，特发性肺纤维化由于肺纤维化快速进展，预后比特发性结节病差。通过包括胰蛋白酶消化产物的质谱指纹分析在内的多种方法，鉴定出38种蛋白质或蛋白质片段，这些蛋白质或片段以前从未在BALF样本中鉴定过。两组患者BALF蛋白质图谱的比较显示，有32个点的相对丰度存在与疾病相关的统计学显著差异。在结节病中，我们发现几种血浆蛋白的量增加，而在特发性肺纤维化中，我们观察到低分子量蛋白有统计学显著增加，其中许多蛋白参与炎症过程（如巨噬细胞移动抑制因子和钙粒蛋白）或抗氧化反应（如抗氧化过氧化物酶体酶和硫氧还蛋白过氧化物酶2）。二维电泳使我们能够鉴定新的BALF蛋白，并对结节病和特发性肺纤维化患者的蛋白质组成进行表征。两种疾病凝胶的比较表明，它们在BALF蛋白质谱方面存在差异，正如它们在免疫反应类型方面存在差异一样。
