Liang Wenzhao, Ward Liam J, Karlsson Helen, Ljunggren Stefan A, Li Wei, Lindahl Mats, Yuan Xi-Ming
Occupational and Environmental Medicine Center, and Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden.
Department of Neurology, China-Japan Union Hospital, Jilin University, Changchun, China.
Sci Rep. 2016 May 20;6:26231. doi: 10.1038/srep26231.
The heterogeneity of atherosclerotic tissue has limited comprehension in proteomic and metabolomic analyses. To elucidate the functional implications, and differences between genders, of atherosclerotic lesion formation we investigated protein profiles from different regions of human carotid atherosclerotic arteries; internal control, fatty streak, plaque shoulder, plaque centre, and fibrous cap. Proteomic analysis was performed using 2-DE with MALDI-TOF, with validation using nLC-MS/MS. Protein mapping of 2-DE identified 52 unique proteins, including 15 previously unmapped proteins, of which 41 proteins were confirmed by nLC-MS/MS analysis. Expression levels of 18 proteins were significantly altered in plaque regions compared to the internal control region. Nine proteins showed site-specific alterations, irrespective of gender, with clear associations to extracellular matrix remodelling. Five proteins display gender-specific alterations with 2-DE, with two alterations validated by nLC-MS/MS. Gender differences in ferritin light chain and transthyretin were validated using both techniques. Validation of immunohistochemistry confirmed significantly higher levels of ferritin in plaques from male patients. Proteomic analysis of different plaque regions has reduced the effects of plaque heterogeneity, and significant differences in protein expression are determined in specific regions and between genders. These proteomes have functional implications in plaque progression and are of importance in understanding gender differences in atherosclerosis.
动脉粥样硬化组织的异质性限制了蛋白质组学和代谢组学分析中的理解。为了阐明动脉粥样硬化病变形成的功能意义以及性别差异，我们研究了人类颈动脉粥样硬化动脉不同区域的蛋白质谱；内部对照、脂纹、斑块肩部、斑块中心和纤维帽。使用二维电泳（2-DE）结合基质辅助激光解吸电离飞行时间质谱（MALDI-TOF）进行蛋白质组分析，并使用纳升液相色谱-串联质谱（nLC-MS/MS）进行验证。二维电泳的蛋白质图谱鉴定出52种独特蛋白质，包括15种先前未映射的蛋白质，其中41种蛋白质经nLC-MS/MS分析确认。与内部对照区域相比，18种蛋白质在斑块区域的表达水平有显著变化。9种蛋白质显示出位点特异性改变，与性别无关，且与细胞外基质重塑有明显关联。5种蛋白质在二维电泳中显示出性别特异性改变，其中2种改变经nLC-MS/MS验证。使用这两种技术均验证了铁蛋白轻链和转甲状腺素蛋白的性别差异。免疫组织化学验证证实男性患者斑块中铁蛋白水平显著更高。对不同斑块区域的蛋白质组分析减少了斑块异质性的影响，并确定了特定区域和性别之间蛋白质表达的显著差异。这些蛋白质组对斑块进展具有功能意义，对于理解动脉粥样硬化中的性别差异很重要。
