Abstract
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.
Highlights
(FPP) and geranylgeranyl pyrophosphate (GGPP), are the precursors for posttranslational protein modifications prenylation[13,14]
A gel-free quantitative proteomic profiling in murine Raw 264.7 macrophage cells along with the treatment of statin and LPS-stimulation allowed to understand the changes of protein expression and proteins activities
We identified 334 differentially expressed proteins and 133 commonly shared proteins with high confidence in LPS and statin treated macrophages
Summary
(FPP) and geranylgeranyl pyrophosphate (GGPP), are the precursors for posttranslational protein modifications prenylation[13,14]. Analysis of biological networks using differentially expressed proteins after the treatment of statin and LPS in Raw 264.7 macrophage cells can predict possible canonical pathways, upstream regulators, and functional metabolic networks. This study presents a discovery based label-free quantitative proteomics due to the combined treatment of LPS and statin on murine Raw 264.7 macrophage cells. Raw 264.7 macrophage cells were stimulated by the proinflammatory mediator LPS as well as drug statin to understand the regulation and mechanism of proteins comprehensively during inflammatory response, and the relative effect of LPS and statin on the protein level. This study included an effective compilation of statin and LPS treated liquid chromatography-mass spectrometry (LC-MS/MS) proteomics data and elucidates their correlation with the anti and proinflammatory responses of immune cells. We narrowed down a significant protein target, plectin by proteomics study and further validated its expression by biochemical and molecular biological methods and provided convincing evidence of its influence due to statin drug and pathogenic stimuli
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
