Interleukin-1 inhibits osmotically induced calcium signaling and volume regulation in articular chondrocytes

Abstract

Articular chondrocytes respond to osmotic stress with transient changes in cell volume and the intracellular concentration of calcium ion ([Ca(2+)](i)). The goal of this study was to examine the hypothesis that interleukin-1 (IL-1), a pro-inflammatory cytokine associated with osteoarthritis, influences osmotically induced Ca(2+) signaling. Fluorescence ratio imaging was used to measure [Ca(2+)](i) and cell volume in response to hypo- or hyper-osmotic stress in isolated porcine chondrocytes, with or without pre-exposure to 10-ng/ml IL-1alpha. Inhibitors of IL-1 (IL-1 receptor antagonist, IL-1Ra), Ca(2+) mobilization (thapsigargin, an inhibitor of Ca-ATPases), and cytoskeletal remodeling (toxin B, an inhibitor of the Rho family of small GTPases) were used to determine the mechanisms involved in increased [Ca(2+)](i), F-actin remodeling, volume adaptation and active volume recovery. In response to osmotic stress, chondrocytes exhibited transient increases in [Ca(2+)](i), generally followed by decaying oscillations. Pre-exposure to IL-1 significantly inhibited regulatory volume decrease (RVD) following hypo-osmotic swelling and reduced the change in cell volume and the time to peak [Ca(2+)](i) in response to hyper-osmotic stress, but did not affect the peak magnitudes of [Ca(2+)](i) in those cells that did respond. Co-treatment with IL-1Ra, thapsigargin, or toxin B restored these responses to control levels. The effects were associated with alterations in F-actin organization. IL-1 alters the normal volumetric and Ca(2+) signaling response of chondrocytes to osmotic stress through mechanisms involving F-actin remodeling via small Rho GTPases. These findings provide further insights into the mechanisms by which IL-1 may interfere with normal physiologic processes in the chondrocyte, such as the adaptation or regulatory responses to mechanical or osmotic loading.