Elsevier

International Immunopharmacology

Volume 39, October 2016, Pages 149-157
International Immunopharmacology

Advanced oxidation protein products induce catabolic effect through oxidant-dependent activation of NF-κ B pathway in human chondrocyte

https://doi.org/10.1016/j.intimp.2016.07.018Get rights and content

Highlights

  • AOPPs increased catabolic factors expression and decreased glycosaminoglycan production.

  • AOPPs-induced catabolic factor expression was mediated by NF-κB activation.

  • NF-κB activation was associated with RAGE-mediated, NADPH-dependent ROS generation pathway.

Abstract

Advanced oxidation protein products (AOPPs) have been shown to participate in the progression of rheumatoid arthritis (RA). However, the effect of AOPPs accumulation on catabolic effect in human chondrocyte and the underlying mechanism(s) remain unclear. The present study demonstrated that AOPPs inhibited cell viability and glycosaminoglycan (GAG) production in human chondrocyte. Exposure of chondrocyte to AOPPs significantly increased the production of catabolic factors, such as cyclooxygenase-2 (COX-2), matrix metalloproteinase 3 (MMPs)-3 and MMP-13. AOPPs stimulation induced ROS generation and NF-κ B p65 phosphorylation, which could be inhibited by soluble receptor for advanced glycan end products (sRAGE), NADPH oxidase inhibitor (apocynin), ROS scavenger (N-acetyl-cysteine, NAC). Furthermore, NF-κ B inhibitor Bay11-7082 significantly reversed the AOPPs-induced expression of catabolic factors and phosphorylation of NF-κ B p65. Targeting AOPPs-triggered catabolic effect might be as a promising option for patients with RA.

Introduction

Rheumatoid arthritis (RA) is a systemic inflammatory disorder characterised by accumulation of pro-inflammatory cytokines in joint cavity, leading to destruction of cartilage and bone tissue of the joints [1]. Recent studies have demonstrated that RA is associated with increased modification of proteins. A typical representation is the formation and accumulation advanced oxidation protein products (AOPPs) [2].

AOPP appear as novel markers of the oxidative stress involved in RA, as well as being vital inflammatory mediators able to amplify neutrophil activation. In fact, AOPPs have been considered as true mediators of the pro-inflammatory effects via Redox-dependent pathway [3], [4], [5], [6]. Our recent studies also demonstrated that exposure to AOPP can cause cartilage destruction in rabbit arthritic model and induce apoptosis in murine chondrocyte [7], [8]. Although emerging data recognize AOPPs as pathogenic factors of RA, the underlying mechanisms have not been investigated.

On the other hands, as inflammation in general, RA is triggered by pro-inflammatory molecules, including enzymes (COX-2 and metalloproteinases) [9], [10]. These catabolic factors mediate the degradation of cartilagineus extracellular matrix (ECM), as shown by the blockade of glycosaminoglycan (GAG) products. It is well documented that induction of NF-κ B activity in chondrocytes would facilitate degradation of the extracellular matrix of cartilage by over-production of those catabolic factors [11], [12], [13], [14]. However, to the best of our knowledge, the mechanisms underlying AOPPs-induced expression of catabolic factors and cartilage destruction have not been completely elucidated.

The present study was designed to test the hypothesis that AOPPs may trigger catabolic effect in human articular chondrocyte. We demonstrated a role of RAGE, ROS and NF-κ B in AOPPs-induced GAG loss and production of catabolic factors in primary human chondrocytes. These results suggest a new mechanism underlying catabolic effect of AOPPs and provided a link between protein oxidative damage and the progression of RA.

Section snippets

Reagents

Buman serum albumin (BSA), collagenase, Limulus Amoebocyte Lysate kit was purchased from Sigma (St Louis, MO, USA). HOCl was purchased from Fluke (Buchs, Switzerland). Fetal bovine serum (FBS) and MTT were purchased from Gibco BRL (Melbourne, Australia). TRI Reagent® and cDNA Synthesis Kit were purchased from Molecular Research Center (Invitrogen, Carlsbad, CA, USA). PrimeScript TM RT reagent kit was from Takara Biotechnology (Dalian, China). GoTaq® Green Master Mix was purchased from Promega

Effects of AOPPs on cellular viability and GAG production in human chondrocyte

To determine whether AOPPs stimulation have toxicity for human chondrocyte, we have measured cell viability and lactate dehydrogenase (LDH) release. As shown in Fig.1, inhibition effects were observed in cells exposed to 200–400 μg/ml of AOPPs for 24 h, in comparison with the control medium and native BSA (Fig. 1a). Besides, cell viability was markedly inhibited for cells treated with 200 μg/ml of AOPPs for 24–72 h (Fig. 1b). In agreement with the results on cell viability, LDH levels were

Discussion

The formation and accumulation of AOPPs were well documented in RA [2]. Our previous study has confirmed that AOPPs could accelerate cartilage degradation in rabbit arthritis models and apoptosis in murine chondrocyte [7], [8]. But the mechanisms of its actions, especially on catabolic effect of human chondrocyte remain unknown. The present study demonstrated that AOPPs significantly enhanced catabolic effect of cultured human chondrocyte through the interaction of RAGE, NADPH-dependent ROS

Conflicts of interest

The authors declare no conflict of interest.

Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No.81272042).

References (29)

  • Q. Wu et al.

    Advanced oxidation protein products induce chondrocyte apoptosis via receptor for advanced glycation end products-mediated, redox-dependent intrinsic apoptosis pathway

    Apoptosis

    (2015)
  • H. Yu et al.

    Effect of advanced oxidation protein products on articular cartilage and synovium in a rabbit osteoarthritis model

    Orthop. Surg.

    (2015)
  • M.P. Vincenti

    The matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) genes. Transcriptional and posttranscriptional regulation, signal transduction and cell-type-specific expression

    Methods Mol. Biol.

    (2001)
  • M.B. Goldring et al.

    The regulation of chondrocyte function by proinflammatory mediators: prostaglandins and nitric oxide

    Clin. Orthop. Relat. Res.

    (2004)
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