Analysis of Expression of Angiogenic and Angiostatic Chemokines and Their Receptors in Synovial Tissue by Quantitative Real-Time PCR

Abstract

Chemokines are generally believed to play an important role in immunopathogenesis of different joint diseases. Synovial fluid is the primary bio-material, along with peripheral blood, for evaluation of patients with joint pathology (rheumatoid arthritis, osteoarthritis, psoriatic arthritis, etc). However, analysis of synovial membranes is also of importance, since the main pathologic processes evolve at this site. Analysis of chemokines and their receptors in synovium may provide more complete information about immunopathogenesis of articular disease of different etiologies. We carried out optimization procedures for real-time reverse-transcription PCR analysis, in order to quantify specific mRNA expression of angiogenic and angiostatic chemokines and their receptors in synovium. All the stages, from sampling the biological material and until analysis of results are described in details. Normal ranges for mRNA expression rates are provided, as based on examination of subjects with previous traumatic joint injury, without any clinical signs of current systemic or local inflammation. We demonstrate that this method can be informative for comparing intra-articular expression of chemokines and their receptors between different clinical groups. It can also be used to investigate the role of chemokines in appropriate immunopathological events.