Functional Insights into the Sphingolipids C1P, S1P, and SPC in Human Fibroblast-like Synoviocytes by Proteomic Analysis

Int J Mol Sci. 2024 Jul 31;25(15):8363. doi: 10.3390/ijms25158363.

Abstract

The (patho)physiological function of the sphingolipids ceramide-1-phosphate (C1P), sphingosine-1-phosphate (S1P), and sphingosylphosphorylcholine (SPC) in articular joints during osteoarthritis (OA) is largely unknown. Therefore, we investigated the influence of these lipids on protein expression by fibroblast-like synoviocytes (FLSs) from OA knees. Cultured human FLSs (n = 7) were treated with 1 of 3 lipid species-C1P, S1P, or SPC-IL-1β, or with vehicle. The expression of individual proteins was determined by tandem mass tag peptide labeling followed by high-resolution electrospray ionization (ESI) mass spectrometry after liquid chromatographic separation (LC-MS/MS/MS). The mRNA levels of selected proteins were analyzed using RT-PCR. The 3sphingolipids were quantified in the SF of 18 OA patients using LC-MS/MS. A total of 4930 proteins were determined using multiplex MS, of which 136, 9, 1, and 0 were regulated both reproducibly and significantly by IL-1β, C1P, S1P, and SPC, respectively. In the presence of IL-1ß, all 3 sphingolipids exerted ancillary effects. Only low SF levels of C1P and SPC were found. In conclusion, the 3 lipid species regulated proteins that have not been described in OA. Our results indicate that charged multivesicular body protein 1b, metal cation symporter ZIP14, glutamine-fructose-6-P transaminase, metallothionein-1F and -2A, ferritin, and prosaposin are particularly interesting proteins due to their potential to affect inflammatory, anabolic, catabolic, and apoptotic mechanisms.

Keywords: ceramide-1-phosphate; functions; inflammation; lysosphingomyelin; mass spectrometry; osteoarthritis; proteomics; sphingolipid; sphingosine-1-phosphate; sphingosylphosphorylcholine.

MeSH terms

  • Aged
  • Cells, Cultured
  • Ceramides* / metabolism
  • Female
  • Fibroblasts* / metabolism
  • Humans
  • Interleukin-1beta / metabolism
  • Lysophospholipids* / metabolism
  • Male
  • Middle Aged
  • Osteoarthritis / genetics
  • Osteoarthritis / metabolism
  • Osteoarthritis / pathology
  • Osteoarthritis, Knee / genetics
  • Osteoarthritis, Knee / metabolism
  • Osteoarthritis, Knee / pathology
  • Phosphorylcholine / analogs & derivatives
  • Proteomics* / methods
  • Sphingolipids / metabolism
  • Sphingosine* / analogs & derivatives
  • Sphingosine* / metabolism
  • Synoviocytes* / metabolism
  • Synoviocytes* / pathology
  • Tandem Mass Spectrometry

Substances

  • Lysophospholipids
  • sphingosine 1-phosphate
  • Sphingosine
  • ceramide 1-phosphate
  • sphingosine phosphorylcholine
  • Ceramides
  • Sphingolipids
  • Interleukin-1beta
  • Phosphorylcholine