Advanced oxidation protein products induce catabolic effect through oxidant-dependent activation of NF-κ B pathway in human chondrocyte
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)
- et al.
The role of advanced oxidation protein products in regulation of dendritic cell function
Free Radic. Biol. Med.
(2002) - et al.
Inhibitory effects of melittin on the production of lipopolysaccharide-induced matrix metalloproteinase 3 in human osteoarthritic chondrocytes
Toxicon
(2007) - et al.
PCB126 induces apoptosis of chondrocytes via ROS-dependent pathways
- et al.
Advanced glycation end products increases matrix metalloproteinase-1, -3, and -13, and TNF-alpha in human osteoarthritic chondrocytes
FEBS Lett.
(2007) - et al.
T-cell regulation in rheumatoid arthritis
Curr. Opin. Rheumatol.
(2004) - et al.
Investigation of protein oxidation and lipid peroxidation in patients with rheumatoid arthritis
Cell Biochem. Funct.
(2006) - et al.
Advanced oxidation protein products as novel mediators of inflammation and monocyte activation in chronic renal failure
J. Immunol.
(1998) - et al.
Advanced oxidation protein products induce inflammatory response in fibroblast-like synoviocytes through NADPH oxidase -dependent activation of NF-kappaB
Cell. Physiol. Biochem.
(2013) - et al.
Advanced oxidation protein products inhibit proliferation and differentiation of rat osteoblast-like cells via NF-kappa B pathway
Cell. Physiol. Biochem.
(2009) - et al.
Oxidant/antioxidant status of plasma samples from patients with rheumatoid arthritis
Rheumatol. Int.
(1999)
Advanced oxidation protein products induce chondrocyte apoptosis via receptor for advanced glycation end products-mediated, redox-dependent intrinsic apoptosis pathway
Apoptosis
Effect of advanced oxidation protein products on articular cartilage and synovium in a rabbit osteoarthritis model
Orthop. Surg.
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.
The regulation of chondrocyte function by proinflammatory mediators: prostaglandins and nitric oxide
Clin. Orthop. Relat. Res.
Cited by (17)
N-acetyl-L-cysteine attenuates oxidative stress-induced bone marrow endothelial cells apoptosis by inhibiting BAX/caspase 3 pathway
2023, Biochemical and Biophysical Research CommunicationsThe role of metabolism in chondrocyte dysfunction and the progression of osteoarthritis
2021, Ageing Research ReviewsCitation Excerpt :By regulating GLUT1 expression, normal chondrocytes can adapt to changes in extracellular glucose concentrations at the cell membrane to maintain normal cartilage development (Qing-Xian et al., 2020). Genetic loss of GLUT1 leads to cartilage dysplasia, while continuous elevated GLUT1 expression can degrade cartilage by increasing glucose uptake and producing excessive AGEs (Baumann et al., 2014; Rasheed et al., 2011; Ye et al., 2016). Chondrocyte GLUT1 expression is significantly increased under hypoxic conditions and by pro-inflammatory cytokines (Peansukmanee et al., 2009; Richardson et al., 2003).
Programming changes in GLUT1 mediated the accumulation of AGEs and matrix degradation in the articular cartilage of female adult rats after prenatal caffeine exposure
2020, Pharmacological ResearchCitation Excerpt :Studies found that the genetic defect of cartilage GLUT1 during the developmental period will cause chondrodysplasia [29], which may lead to abnormalities in the structure and joint mechanics of adult articular cartilage to induce OA [13,64]. On the other hand, overexpression of GLUT1 can increase glucose uptake and lead to accumulation of AGEs, which causes cartilage impairment [29–31]. Studies showed that myocardial GLUT1 protein expression is upregulated in IUGR fetuses [65].
Advanced oxidation protein products increase TNF-α and IL-1β expression in chondrocytes via NADPH oxidase 4 and accelerate cartilage degeneration in osteoarthritis progression
2020, Redox BiologyCitation Excerpt :Our previous work demonstrates that intra-articular injections of AOPPs (exogenous injection) accelerate regression of cartilage in rabbit OA models [14]. We also confirmed that AOPPs were able to activate the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-mediated oxidative stress pathway to induce apoptosis of chondrocytes and upregulate the MMP-13 expression [15,16]. The pathogenic role and mechanism of AOPPs in OA disease need further research on the basis of previous extensive efforts into the complex disease before they can be completely elucidated.
Redox and NF-κB signaling in osteoarthritis
2019, Free Radical Biology and MedicineCitation Excerpt :Advanced oxidation protein products (AOPPs) are dityrosine-containing and cross-linking protein products generated by the reaction of proteins with ROS. In human chondrocytes, NADPH oxidase-dependent ROS generation, in combination with the activation of RAGE, mediate AOPPs-induced chondrocyte apoptosis through the activation of NF-κB [213]. Data suggest that increased levels of NO promote joint inflammation by enhancing the production of inflammatory mediators, such as IL-1β and TNF-α, which rapidly activate NF-κΒ.
- 1
Contributed equally to this work.