Interleukin-1 Differentially Modulates Chondrocyte Expression of Cyclooxygenase-2 and Phospholipase A2

Abstract

Interleukin-1β (IL-1) increases the synthesis of prostaglandins as well as the expression of synovial fluid phospholipase A2 (PLA2) mRNA and activity by chondrocytes. In order to examine the potential involvement of cyclooxygense in the induction of prostaglandins by IL-1, the effect of IL-1 on rabbit articular chondrocyte expression of cyclooxygenase enzymes was investigated. By Northern analyses, mRNA for cyclooxygenase-2 (COX-2) was found to be constitutively expressed, and its expression was increased in cultures treated for 24 h with IL-1 (100 ng/ml). Cyclooxygenase-1 mRNA was not detected in control or IL-1-treated cultures. IL-1 caused a concentration-dependent increase in steady-state levels of COX-2 message as assessed by slot-blot analyses. Half-maximal induction of the COX-2 message levels was estimated to require 1.2 ng/ml IL-1. In duplicate slot blots probed with a cDNA for synovial fluid PLA2, half-maximal induction of PLA2 mRNA was estimated to require 0.15 ng/ml IL-1 approximately 10-fold less IL-1 than required for COX-2. In additon, maximal increase in COX-2 message levels was only 3-fold as compared with a 13-fold induction of PLA2. No change in message levels for the intracellular protein, actin, was found between control and experimental cultures. Cycloheximide enhanced COX-2 transcript levels, but did not modulate IL-1 induction of COX-2. Actinomycin D did not inhibit IL-1 augmentation of COX-2 mRNA levels; whereas, induction of PLA2 mRNA levels was completely inhibited, implicating transcriptional mechanisms in the induction of PLA2 but not COX-2 mRNA. These data demonstrate that chondrocyte expression of COX-2 and synovial fluid PLA2 are differentially modulated by IL-1.