Anticoagulants affect matrix metalloproteinase 9 levels in blood samples of stroke patients and healthy controls

Clin Biochem. 2012 Apr;45(6):483-9. doi: 10.1016/j.clinbiochem.2012.01.028. Epub 2012 Feb 8.

Abstract

Objectives: Matrix metalloproteinase-9 (MMP-9) represents a promising marker for acute stroke management. In clinical studies MMP-9 has been quantified by ELISA using differing protocols. We aimed to establish a valid protocol by evaluation of preanalytics.

Design and methods: Blood from stroke patients (n=28) and healthy controls (n=28) was drawn into tubes containing different anticoagulants (EDTA, citrate, lithium-heparin (heparin) and heparin with proteinase inhibitors) and processed after 0, 60 and 240 min. MMP-9 plasma protein and mRNA from mononuclear leukocytes were determined.

Results: In regard to anticoagulants used, samples showed different MMP-9 protein baseline values and kinetics. Stable MMP-9 protein concentrations were only measured from EDTA samples. Particularly in samples with proteinase inhibitors protein and mRNA concentrations increased over time. Kinetics did not differ between patients and controls.

Conclusions: Preanalytics plays a key role for determination of MMP-9. EDTA seems to be a valid anticoagulant for MMP-9 protein measurement.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Anticoagulants / pharmacology
  • Artifacts*
  • Blood Chemical Analysis / methods*
  • Case-Control Studies
  • Citric Acid / pharmacology*
  • Enzyme Stability
  • Female
  • Heparin / pharmacology*
  • Humans
  • Leukocytes, Mononuclear / enzymology
  • Leukocytes, Mononuclear / metabolism
  • Male
  • Matrix Metalloproteinase 9 / blood*
  • Matrix Metalloproteinase 9 / genetics
  • Middle Aged
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Statistics, Nonparametric
  • Stroke / blood*
  • Stroke / enzymology
  • Transcription, Genetic

Substances

  • Anticoagulants
  • RNA, Messenger
  • Citric Acid
  • Heparin
  • Matrix Metalloproteinase 9