Protein profile in neuroblastoma cells incubated with S‐ and R‐enantiomers of ibuprofen by iTRAQ‐coupled 2‐D LC‐MS/MS analysis: Possible action of induced proteins on Alzheimer's disease

Abstract

Ibuprofen is a member of the proprionic acid group of nonsteroidal anti-inflammatory drugs (NSAID), with the S-enantiomer being more active than the R-enantiomer. It has been shown to display protective effects against neuroinflammation, which is linked to the pathogenesis of several neurodegenerative disorders, including Alzheimer's disease (AD). While its prophylactic effect on AD has been suggested, a comprehensive understanding of its mechanism of action remains unclear. Using iTRAQ-coupled 2-D LC-MS/MS analysis, we report here the first study of protein profiles of neuroblastoma cells incubated separately with the two enantiomers of ibuprofen. Three types of cellular proteins, including metabolic enzymes, signaling molecules and cytoskeletal proteins, displayed changes. The changes in the level of a number of enzymes involved in fatty acid synthesis and antioxidant activity in cells incubated with the S-enantiomer were further supported by the real-time PCR analysis as well as the reduced level of reactive oxygen species in cells incubated with the S-enantiomer of ibuprofen. Our findings, therefore, provide the possible mechanism of ibuprofen-induced proteins on AD, and the beneficial effects of ibuprofen in reducing the development of AD.

iTRAQ-coupled 2-D LC–MS/MS analysis of cytoplasmic protein profile in Escherichia coli incubated with apidaecin IB

“…iTRAQ-labeled peptide mixtures were analyzed as described previously with a slight modification [19][20][21][22][23].The analysis was performed on combination of an Agilent 1200 nanoflow LC system (Agilent Technologies Inc., USA) and a 6530 Q-TOF mass spectrometer (Agilent Technologies Inc., USA). In the first dimension 3 μl of the combined peptide mixture was loaded onto the PolySulfoethyl A SCX column (0.3 × 50 mm, 5 μm) and was eluted stepwise by injecting salt plugs of 10 different molar concentrations of 10, 20, 30, 40, 50, 60, 80,100, 300, and 500 mM KCl solution.…”

Protein expression profiles in osteoblasts in response to differentially shaped hydroxyapatite nanoparticles

“…The quantization of RNA was determined by measuring the absorbance at 260 nm (A 260 ) in a spectrophotometer. To accurately quantify the original amount of target mRNAs in samples, Real-time RT-PCR was applied using iSCRIPT one-step RT-PCR kit (Bio-Rad) as described [13]. The real-time PCR was carried out in an IQ 5 multicolor Real-time PCR detection system (Bio-Rad).…”

“…Total RNA was extracted from osteoblast cells in the presence of soluble nano HA powders at the initial culturing stage of 4 days using RNeasy mini kit (Qiagen) as described [13]. The flow-out RNA sample after centrifuged was added DNase to remove genomic DNA contamination.…”

VKORC1 Pharmacogenetics and Pharmacoproteomics in Patients on Warfarin Anticoagulant Therapy: Transthyretin Precursor as a Potential Biomarker

“…Peptide identifications and quantization was performed using Proteinpilot software packages (Applied Biosystems). Each MS/MS spectrum was blasted against the human protein database and protein identifications were accepted based on Proteinpilot confidence scores, which are based on peptide scores, number of peptides and distance from nearest neighbor [26], [27]. Relative quantification of proteins in the case of iTRAQ was performed on the MS/MS scans and is the ratio of the areas under the peaks at 114, 115, 116, and 117 Da which represents the masses of the tags that correspond to the iTRAQ reagents.…”