Proteomic Response of Human Umbilical Vein Endothelial Cells to Histamine Stimulation

Abstract

The histamine receptors (HRs) represent a subclass of G protein-coupled receptors (GPCRs) and comprise four subtypes. Due to their numerous physiological and pathological effects, HRs are popular drug targets for the treatment of allergic reactions or the regulation of gastric acid secretion. Hence, an understanding of the functional selectivity of HR ligands has gained importance. These ligands can bind to specific GPCRs and selectively activate defined pathways. Supporting the activation of a therapeutically necessary pathway without the activation of other signaling cascades can result in drugs with more specific activity and fewer side effects. To evaluate the cellular consequences resulting from receptor binding, comprehensive analyses of cellular protein alterations upon incubation with ligands are required. For this purpose, endothelial cells are treated with histamine, as the endogenous ligand of HRs, to obtain a global overview of its cellular effects. Quantitative proteomics and pathway analyses of histamine-treated and untreated cells reveal enrichment of the nuclear factor-κB and tumor necrosis factor signaling pathways, cytokine-cytokine receptor interactions, complement and coagulation cascades, and acute inflammatory processes upon histamine treatment. This strategy offers the opportunity to monitor HR-mediated signaling in a multidimensional manner.

Functionally selective activation of the dopamine receptor D2 is mirrored by the protein expression profiles

“…Lastly, the samples were centrifuged for 10 min at 12,000 × g and 4 °C. The dried supernatants were subjected to C18 purification as described previously with minor modifications 15 . Briefly, conventional 200 µL-pipette tips were packed with five layers of 3 M Empore disk C18 material.…”

“…In contrast, the analysis of cellular protein expression at later stages focuses on the effects induced by GPCR signaling. Thus, Emirbayer et al revealed an enrichment of cellular effects such as cytokine–cytokine receptor interactions and acute inflammatory processes in histamine-treated endothelial cells after 24 h 15 . Dong et al used proteome analysis to investigate cellular effects induced by the stimulation of the orphan GPCR bombesin-like receptor 3 for one to twelve hours, indicating a regulation of cell death and protein synthesis, particularly mRNA translation 16 .…”

The complex functioning of the complement system in xenotransplantation

“…Some factors may dysregulate the complement and coagulation systems, activate platelets, and result in consumptive coagulopathy, which is a main manifestation hindering xenograft survival . For example, (a) after adhering to activated platelets, pentameric C‐RP complex dissociates into monomeric C‐RP and recruits more platelets, and activates the AP; (b) antibodies against platelet membrane glycoproteins may activate the CP through C1q; (c) antibodies against CD55 and CD59 on platelets may interfere with complement regulation and cause exacerbated injury to platelets and immune thrombocytopenia; (d) after being stimulated by histamine, human umbilical vein endothelial cells may become enriched in local acute inflammatory molecules, including molecules involved in both the complement and coagulation cascades; (v) dysregulation of the complement system and drug‐induced injury, for example, by sirolimus, cyclosporine, tacrolimus, may decrease expression of vascular endothelial growth factor in a graft, resulting in endothelial injury and complement activation, leading to thrombotic microangiopathy and consumptive coagulopathy.…”

Analysis of Differentially Expressed Proteins and Modifications Induced by Formaldehyde Using LC-MS/MS

“…To understand the molecular mechanism of FA-induced damage to biological macromolecules, a variety of methods have been employed to determine the levels and effects of FA, including spectrophotometry, fluorescence, liquid chromatography (LC) and mass spectrometry (MS) [16][17][18][19][20][21]. At present, liquid chromatography tandem mass spectrometry (LC-MS/MS) is a powerful tool for proteomics analysis and protein modification identification [22,23], and thus can provide more detailed information regarding protein alterations induced by FA compared to traditional methods [24][25][26][27]. However, the effect of FA at the protein level remains to be explored.…”