Abstract：
Occupational silica exposure caused a serious disease burden of silicosis. There is currently a lack of sensitive and effective biomarkers for silicosis, and the pathogenesis of silicosis is unclear. Exosomes were significant in the pathogenesis of silicosis, and our study was carried out from exosomal proteomics and cytokine analysis. Firstly, the plasma levels of cytokines were detected using a Luminex multiplex assay, and the results indicated that the plasma levels of TNF-α, IL-6, CCL2, CXCL10, and PDGF-AB were significantly higher in silicosis patients than in silica-exposed workers and controls (p<0.05). After correlation analysis, the plasma levels of cytokines were positively correlated with exosomal protein concentration. Secondly, data-independent acquisition (DIA) was performed on plasma-derived exosomes in the screening population, which identified 88, 151, 293, and 53 differentially expressed proteins (DEPs) in exposure/control, silicosis/control, silicosis/exposure, and silicosis stage Ⅲ/silicosis stage Ⅰ groups respectively. After parallel reaction monitoring (PRM) in an independent verification population, the results indicated that the changing trend of 15 DEPs was coincident in screening and verification results. The result of correlation analysis indicated that the plasma level of TNF-α was negatively correlated with the expression of exosomal DSP, KRT78, SERPINB12, and CALML5. The AUC of combined determination of TNF-α and CALML5 reached 0.900, with a sensitivity of 0.714 and a specificity of 0.933. Overall, our study revealed the exosomal proteomic profiling of silicosis patients, silica-exposed workers, and controls, indicating that exosomes were significant in the pathogenesis of silicosis. It also revealed that the combined of the plasma levels of cytokines and the expression of exosomal DEPs could increase determination efficiency. This study provided directions for the development of silicosis biomarkers and a scientific basis for the pathogenesis research of silicosis in the future.
摘要：
职业性硅暴露导致矽肺严重的疾病负担。目前对矽肺病缺乏敏感有效的生物标志物,矽肺的发病机制尚不清楚。外泌体在矽肺的发病机制中具有重要意义，我们的研究是从外泌体蛋白质组学和细胞因子分析进行的。首先,使用Luminex多重测定法检测细胞因子的血浆水平，结果表明，血浆中的TNF-α水平，矽肺患者的IL-6、CCL2、CXCL10和PDGF-AB显著高于二氧化硅暴露工人和对照组(p<0.05)。经过相关性分析,血浆细胞因子水平与外泌体蛋白浓度呈正相关。其次,对筛查人群中的血浆来源的外来体进行数据独立采集（DIA），在暴露/对照中鉴定出88、151、293和53种差异表达蛋白（DEP），矽肺/控制，矽肺/暴露，分别为矽肺Ⅲ期/矽肺Ⅰ期。在独立验证群体中进行平行反应监测(PRM)后，结果表明，15个DEP的变化趋势在筛查和验证结果上是一致的。相关性分析结果显示,血浆TNF-α水平与外泌体DSP的表达呈负相关,KRT78、SERPINB12和CALML5。联合测定TNF-α和CALML5的AUC达到0.900，灵敏度为0.714，特异性为0.933。总的来说,我们的研究揭示了矽肺患者的外泌体蛋白质组学分析，接触二氧化硅的工人，和控制，表明外泌体在矽肺的发病机制中具有重要意义。还揭示了细胞因子的血浆水平和外泌体DEP的表达的组合可以提高测定效率。本研究为矽肺生物标志物的开发提供了方向,为今后矽肺的发病机制研究提供了科学依据。
