Dataset Information

C2C12 PEA and Ibuprofen LC-MS/MS

ABSTRACT: This study investigated the effects of palmitoylethanolamide (PEA) and ibuprofen (IBU) treatment on the dynamic regulation of the skeletal muscle proteome using a C2C12 myotube model. Differentiated myotubes were treated for 36 hours with vehicle control (VC), PEA (10 µM), or IBU (100 µM). Dynamic proteome profiling was performed using deuterium oxide (D₂O) metabolic labelling combined with high-resolution LC–MS/MS analysis on an Orbitrap Q-Exactive (QE) instrument. Samples were collected at 0 h, 12 h, 24 h, and 36 h, with n = 3 biological replicates per timepoint and condition. Protein-specific fractional synthesis rates (FSR) were derived from the 12–36 h labelling period, and relative protein abundance was quantified at 36 h.

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture

SUBMITTER: Connor Stead

LAB HEAD: Dan Owens

PROVIDER: PXD069882 | Pride | 2026-05-25

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	C2C12_PEA.MSMS.mgf	Mgf
	C2C12_PEA.mzid	Mzid
	C2C12_PEA_ABSOLUTE_protein_measurements.csv	Csv
	C2C12_PEA_RELATIVE_protein_measurements.csv	Csv
	C2C12_PEA_peptide_measurements.csv	Csv

Items per page:

1 - 5 of 38

Publications

The Effects of Palmitoylethanolamide or Ibuprofen on the Abundance Profile and Synthesis Rate of Proteins in C2C12 Skeletal Myotubes.

Cole Paige L PL Stead Connor A CA Burniston Jatin G JG Owens Daniel J DJ

FASEB bioAdvances 20260423 4

Palmitoylethanolamide (PEA) and ibuprofen (IBU) exert anti-inflammatory effects that may influence skeletal muscle adaptation; however, their impact on muscle proteome dynamics remains unclear. Dynamic proteome profiling was performed in differentiated C2C12 myotubes treated for 36 h with D<sub>2</sub>O and either vehicle control (VC), PEA (10 μM), or IBU (100 μM). Protein-specific fractional synthesis rates (FSR; 1541 proteins) and relative protein abundances at 36 h (3085 proteins) were quanti ...[more]

PMID: 42038098

Similar Datasets

Project description:Palmitoylethanolamide (PEA) is an endogenous lipid mediator with recognized immunomodulatory actions, yet its effects in skeletal muscle remain poorly defined. We investigated whether PEA influences myogenesis and profiled the acute transcriptomic response of differentiated C2C12 myotubes. Dose-tolerability assays identified reduced metabolic activity and viability at 100 µM PEA, so 10 µM was used for subsequent experiments. Across differentiation, PEA decreased myotube number (90.3 ± 10.6 vs 112.6 ± 10.1 in control) while increasing nuclear fusion index (37.8 ± 5.7% vs 30.7 ± 3.2%); myotube area was unchanged. In myoblasts, 24 h PEA exposure increased the proportion of cells in G0/G1 (48.2 ± 1.2% vs 42.3 ± 1.9%) with a concomitant reduction in S-phase (21.7 ± 1.2% vs 25.5 ± 1.2%), consistent with G1 arrest and reduced proliferative drive. RNA sequencing revealed a distinct transcriptional signature after 24 h PEA treatment, with 1,952 differentially expressed genes (1,028 upregulated; 924 downregulated, q < 0.05). Pathway-level changes encompassed cytokine–receptor interactions, chemokine, JAK–STAT, NF-κB, NOD-like and Toll-like receptor signalling. PEA downregulated NF-κB target/pro-inflammatory cytokine genes (Il1a, Il1b, Il12a, Il33, Tnfrsf19) while upregulating interferon-related and chemokine genes (e.g., Stat2, Oas1b, Gbp7, Cxcl14, Ccl5), indicating an anti-inflammatory/immune-priming profile. Notably, N-acylethanolamine acid amidase (Naaa), a lysosomal PEA hydrolase, was highly expressed and significantly induced, whereas fatty acid amide hydrolase (Faah) was low and unchanged, suggesting a muscle-specific bias toward NAAA-dependent PEA metabolism. Peroxisome proliferator-activated receptor-α (PPARα) was expressed at low levels and unaffected by PEA, indicating that the observed responses occur largely independent of canonical PPARα signalling. Together, these data show that PEA biases skeletal muscle toward a less proliferative but more fused and inflammation-resolving phenotype, accompanied by transcriptional reprogramming of immune pathways and preferential engagement of NAAA. While in vitro concentrations exceed reported plasma levels and mRNA changes may not fully predict protein/function, these findings provide the first comprehensive transcriptomic characterisation of PEA in myotubes and motivate in vivo studies to test whether PEA’s dual actions translate into benefits for muscle regeneration, adaptation, or anti-atrophy interventions.