Department of Chemistry and Biochemistry, University of Texas at Arlington, TX, 76019, USA.
Department of Pathology, UT Southwestern Medical Center, Dallas, TX, 75390, USA.
Sci Rep. 2018 Jan 9;8(1):164. doi: 10.1038/s41598-017-18533-1.
A significant component of immune biology research is the investigation of protein encoding genes that play central roles in contributing inflammatory response. A gel-free quantitative bottom-up proteomics study was performed on immune cell macrophages after the combined treatment of lipopolysaccharide (LPS) and statin drugs using mass spectrometry and a detailed bioinformatics analyses were conducted. Systematic bioinformatics analysis was applied for discovering novel relationships among proteins and effects of statin and lipopolysaccharide in macrophage cells. Based on gene ontology, majority of protein encoding genes was involved in metabolic and cellular processes and are actively associated with binding, structural molecular, and catalytic activity. Notably, proteomic data analyzed by Ingenuity Pathway Analysis (IPA), discovered the plectin and prohibitin 2 protein interactions network and inflammatory-disease based protein networks. Two up-regulated proteins, plectin and prohibitin 2, were further validated by immunoblotting. Plectin was also cross-validated by immunocytochemistry, since its expression was highly modulated by statin but inhibited during LPS-stimulation. Collectively, the significant up-regulation of plectin due to the treatment of statin, suggests that statin has a significant impact on the cytoskeletal networks of cells. Plectin might have a significant role in the intermediate filament assembly and dynamics, and possibly stabilizing and crosslinking intermediate filament networks.
免疫生物学研究的一个重要组成部分是研究在炎症反应中起核心作用的蛋白质编码基因。使用质谱法对脂多糖 (LPS) 和他汀类药物联合治疗后的免疫细胞巨噬细胞进行了无胶定量的向下蛋白质组学研究，并进行了详细的生物信息学分析。系统的生物信息学分析用于发现蛋白质之间的新关系以及他汀类药物和脂多糖对巨噬细胞的影响。基于基因本体论，大多数蛋白质编码基因参与代谢和细胞过程，并与结合、结构分子和催化活性密切相关。值得注意的是，通过 IPA（Ingenuity Pathway Analysis）分析蛋白质组学数据，发现了网蛋白和抑制素 2 蛋白相互作用网络以及基于炎症性疾病的蛋白质网络。通过免疫印迹进一步验证了两个上调的蛋白质，网蛋白和抑制素 2。由于他汀类药物的治疗，网蛋白的表达高度调节，而在 LPS 刺激时受到抑制，因此通过免疫细胞化学进行了交叉验证。总的来说，由于他汀类药物的治疗，网蛋白的显著上调表明他汀类药物对细胞的细胞骨架网络有重大影响。网蛋白可能在中间丝组装和动力学中具有重要作用，并且可能稳定和交联中间丝网络。
