Quantitative proteomic analysis of a genetically induced prostate inflammation mouse model via custom 4-plex DiLeu isobaric labeling

Am J Physiol Renal Physiol. 2019 Jun 1;316(6):F1236-F1243. doi: 10.1152/ajprenal.00387.2018. Epub 2019 Apr 17.


Inflammation is involved in many prostate pathologies including infection, benign prostatic hyperplasia, and prostate cancer. Preclinical models are critical to our understanding of disease mechanisms, yet few models are genetically tractable. Here, we present a comparative quantitative proteomic analysis of urine from mice with and without prostate-specific inflammation induced by conditional prostate epithelial IL-1β expression. Relative quantification and sample multiplexing was achieved using custom 4-plex N,N-dimethyl leucine (DiLeu) isobaric tags and nanoflow ultrahigh-performance liquid chromatography coupled to high-resolution tandem mass spectrometry. Each set of 4-plex DiLeu reagents allows four urine samples to be analyzed simultaneously, providing high-throughput and accurate quantification of urinary proteins. Proteins involved in the acute phase response, including haptoglobin, inter-α-trypsin inhibitor, and α1-antitrypsin 1-1, were differentially represented in the urine of mice with prostate inflammation. Mass spectrometry-based quantitative urinary proteomics represents a promising bioanalytical strategy for biomarker discovery and the elucidation of molecular mechanisms in urological research.

Keywords: benign prostatic hyperplasia; inflammation; interleukin-1β; lower urinary tract symptoms, mass spectrometry; urine proteomics.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Biomarkers / urine
  • Chromatography, High Pressure Liquid
  • Disease Models, Animal
  • High-Throughput Screening Assays
  • Inflammation Mediators / urine*
  • Isotope Labeling*
  • Leucine / analogs & derivatives
  • Leucine / chemistry*
  • Male
  • Mice, Transgenic
  • Prostate / metabolism*
  • Prostate / pathology
  • Prostatitis / genetics
  • Prostatitis / pathology
  • Prostatitis / urine*
  • Proteome*
  • Proteomics / methods*
  • Tandem Mass Spectrometry
  • Time Factors
  • Urinalysis
  • Workflow


  • Biomarkers
  • Inflammation Mediators
  • Proteome
  • Leucine