Interleukin-1β induction of c-fos and collagenase expression in articular chondrocytes: Involvement of reactive oxygen species

Abstract

Interleukin-1 beta (IL-1) is implicated in cartilage destruction in arthritis through promotion of matrix metalloproteinase production. Upregulation of collagenase gene expression by IL-1 is known to require the transactivators Fos and Jun. Recently, reactive oxygen species (ROS) have been suggested to act as intracellular signaling molecules mediating the biological effects of cytokines. Here, we demonstrated ROS production by IL-1-stimulated bovine chondrocytes and that neutralizing ROS activity by the potent antioxidant, N-acetylcysteine, or inhibiting endogenous ROS production by diphenyleneiodonium (DPI), significantly attenuated IL-1-induced c-fos and collagenase gene expression. The inhibitory effect of DPI implicates enzymes such as NADPH oxidase in the endogenous production of ROS. Chondrocytes were also found to produce nitric oxide (NO) upon IL-1 stimulation. That NO may mediate part of the inducing effects of IL-1 was supported by the observation that L-NG-monomethylarginine, a NO synthase inhibitor, partially inhibited IL-1-regulated collagenase expression. Moreover, treatment of chondrocytes with the NO-producing agent, S-nitroso-N-acetylpenicillamine, was sufficient to induce collagenase mRNA levels. In summary, our results suggest that ROS released in response to IL-1 may function as second messengers transducing extracellular stimuli to their targets in the nucleus, leading to augmentation of gene expression.

Heme oxygenase-1 induction modulates microsomal prostaglandin E synthase-1 expression and prostaglandin E2 production in osteoarthritic chondrocytes

“…Pro-inflammatory cytokines such as IL-1 induce the formation of intracellular oxidants which may function as second messengers leading to augmentation of gene expression of degradative enzymes [32]. In addition, ROS have been implicated in cartilage degeneration in OA through the initiation of chondrocyte apoptosis [33], decreased A c c e p t e d M a n u s c r i p t 15 replicative potential, catabolic changes in matrix cartilage, telomere instability and senescence [8].…”

Interleukin-1β stimulates IL-8 expression through MAP kinase and ROS signaling in human gastric carcinoma cells

“…In previous studies, IL-1 was found to stimulate phospholipase A 2 , promoting release of arachidonic acid. Since arachidonic acid can activate NAPDH oxidase to produce superoxide, it is possible that this fatty acid may serve as an intermediate in the IL-1-induced activation of enzymes, leading to the production of ROS (Lo et al, 1998). The observations that NADPH oxidase inhibitors suppressed the IL-1b-and TNF-a-induced ROS production also indicated the involvement of NADPH oxidase in the generation of ROS induced by cytokines (Thannickal and Fanburg, 1995).…”

Role of glucose as a modulator of anabolic and catabolic gene expression in normal and osteoarthritic human chondrocytes

“…The oxidative stress induced by high glucose in OA and also in normal chondrocytes, although for significantly shorter periods in the later [Rosa et al, 2009], may impair normal TGF signaling and consequent responses. Indeed, increased ROS production has been shown to induce the gene expression of various MMPs in chondrocytes from different species [Lo et al, 1998;Gurjar et al, 2001]. Thus, it is possible that high glucose-induced ROS, especially in OA chondrocytes, overcome the inhibitory response induced by TGF, so that MMP-13 gene expression takes longer to be downregulated by TGF when the cells are cultured under high glucose.…”

“…Moreover, that study also showed that OA chondrocytes exposed to a high, hyperglycemia-like extracellular glucose concentration (30 mM) were unable to reduce glucose transport and GLUT-1 content which led to increased glucose accumulation and prolonged Reactive Oxygen Species (ROS) production [Rosa et al, 2009]. Increased oxidative stress has been pointed out as a major pathogenic mechanism of OA [Maneiro et al, 2003;Grishko et al, 2009], and at the cellular level, increased ROS production has been shown to mediate many catabolic responses in chondrocytes [Lo et al, 1998; Mendes et al, 2003a,b].…”

“…Moreover, that study also showed that OA chondrocytes exposed to a high, hyperglycemia-like extracellular glucose concentration (30 mM) were unable to reduce glucose transport and GLUT-1 content which led to increased glucose accumulation and prolonged Reactive Oxygen Species (ROS) production [Rosa et al, 2009]. Increased oxidative stress has been pointed out as a major pathogenic mechanism of OA [Maneiro et al, 2003;Grishko et al, 2009], and at the cellular level, increased ROS production has been shown to mediate many catabolic responses in chondrocytes [Lo et al, 1998; Mendes et al, 2003a,b].Thus, this study was undertaken to determine whether exposure to a high extracellular glucose concentration can shift normal and OA chondrocyte functions towards a pro-catabolic and/or antianabolic gene expression profile, either direct or indirectly by modifying the responses induced by pro-anabolic stimuli, namely TGF.For this purpose, we examined by real time RT-PCR (qRT-PCR) the expression of genes that are important for cartilage homeostasis and OA pathogenesis and that in other cell types have been reported to be modulated by exposure to high glucose [Death et al, 2003;Ho et al, 2007;Andreea et al, 2008;Kim et al, 2008]. Those genes include the MMPs-1 and -13 due to their major role in cartilage matrix degradation, and collagen type II, TIMP-1 and TIMP-2 as important anabolic and anti-catabolic genes, respectively.…”