Effects of heme oxygenase‐1 on bacterial antigen‐induced articular chondrocyte catabolism in vitro

Abstract

This study tested the hypothesis that heme oxygenase-1 (HO-1) expression counteracts bacterial antigen-induced catabolic metabolism in human articular chondrocytes. HO-1 expression was induced in chondrocytes by the iron-containing porphoryin, hemin. Anti-catabolic and anti-apoptotic effects of HO-1 expression were evaluated following bacterial antigen (lipopolysaccharides, LPS) activation of chondrocytes by quantification of cytokine and cartilage matrix protein expression. Effects of HO-1 over-expression on chondrocyte matrix metabolism were evaluated using plasmid-driven protein synthesis. Hemin increased HO-1 expression and LPS increased interleukin-1beta and interleukin-6 gene and protein expression in chondrocytes. Hemin-induced HO-1 decreased LPSinduced interleukin-1beta and interleukin-6 gene and protein expression. Increased HO-1 expression partially reversed LPSsuppression of aggrecan and type II collagen gene expression and suppressed LPS-induced gene expression of IL-6, inducible nitric oxide synthase (iNOS), matrix metalloproteinases (MMPs), and IL-1beta. HO-1 induction was inversely correlated with LPS-induced chondrocyte apoptosis. HO-1 over-expression in chondrocytes decreased matrix protein gene expression. With LPS activation, increased HO-1 expression decreased chondrocyte catabolism, partially reversed LPS-dependent inhibition of cartilage matrix protein expression and protected against apoptosis. Without LPS, hemin-induced HO-1 and plasmid-based over-expression of HO-1 inhibited cartilage matrix gene expression. The results suggest that elevated HO-1 expression in chondrocytes is protective of cartilage in inflamed joints but may otherwise suppress matrix turn over. ß 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31:1943Res 31: -1949Res 31: , 2013 Keywords: heme oxygenase-1; articular chondrocytes; catabolism; bacterial antigen; cartilage matrix proteins Heme oxygenase-1 (HO-1) and heme oxygenase-2 (HO-2) are heme degrading enzymes expressed in mammalian species. 1,2 Although breakdown of heme occurs by similar mechanisms, in many tissues HO-1 is highly inducible isoform whereas HO-2 is constitutively expressed. 3,4 Two of the three heme oxygenase products, biliverdin, and carbon monoxide, modulate immune function and influence tissue metabolism and the third, free iron, is captured by ferritin for reuse. 5,6 Increases in HO-1 occur as in response to tissue stressors, including heme, reactive oxygen species (ROS), unbalanced redox conditions and various inflammatory agents, such as bacterial antigens and pro-inflammatory cytokines. 7,8 In acute or chronic pathogenic states, induction of HO-1 provides protective effects on cells and tissues. 9,10 Chondrocytes express the inducible isoform of HO-1 in osteoarthritic (OA) cartilage explants and in isolated OA chondrocytes. 11,12 In animal models, increased levels of HO-1 are protective of joint tissues for both adjuvant and antigen-induced arthritis. [13][14][15] In OA cartilage explants, HO-1 ind...

Novel anti-inflammatory and chondroprotective effects of the human melanocortin MC1 receptor agonist BMS-470539 dihydrochloride and human melanocortin MC3 receptor agonist PG-990 on lipopolysaccharide activated chondrocytes

“…Numerous studies have highlighted the potential for development of melanocortin peptides for modulating the inflammatory response, including the non-selective pan agonists α-MSH and NDP-α-MSH displaying anti-inflammatory, anti-pyretic effects (Ahmed et al, 2013;Patruno et al, 2018), whilst, more selective compounds [DTrp 8 ]-γ-MSH (Kaneva et al, 2012;Patruno et al, 2018), BMS-470539 dihydrochloride (Leoni et al, 2010;Holloway et al, 2015) and AP214 (Montero-Melendez et al, 2011) modulate inflammatory responses in primary chondrocytes (Capsoni et al, 2015), experimental inflammatory arthritis (Patel et al, 2010), and ischaemic reperfusion injury (Holloway et al, 2015), exerting their effects via human melanocortin MC1 and MC3 receptors via adenylate cyclase as demonstrated here and in agreement with Kaneva et al, 2012 for α-MSH and [DTrp 8 ]-γ-MSH, whilst inhibiting proinflammatory, tissue destructive mediators and inducing pro-resolving anti-inflammatory proteins (including IL-10) (Kaneva et al, 2012), HO-1 (Mawatari et al, 2013) and clearance of apoptotic neutrophils (Patruno et al, 2018). In spite of the potential of melanocortin peptides in modulating inflammation elicited by LPS (Sun et al, 2017), no studies to date have evaluated their effect in LPS (0.1 µg/ml) activated chondrocytes.…”

“…Here, we determined whether these peptides could induce HO-1, previously shown to be upregulated by ACTH, MTII (Lam et al, 2005b) and α-MSH in cardiovascular tissue (Vecsernyes et al, 2017) and a model of retinal damage (Rossi et al, 2016). LPS reduced the expression of the anabolic protein HO-1, as previously seen (Mawatari et al, 2013;Kaneva et al, 2014) and supports a plausible explanation of why elevations in the catabolic proteins IL-1β, IL-6, IL-8, MMP-1, MMP-3 and MMP-13 occur.…”

“…Following determination of the peptides chondroprotective effects, the peptides in the presence of LPS was evaluated against OA markers. LPS stimulation of C-20/A4 chondrocytes led to increases in IL-1β, IL-6, IL-8, MMP-1, MMP-3 and MMP-13 as previously seen in rat (Chen et al, 2013;Hu et al, 2018) and human chondrocytes (Mawatari et al, 2013;Ni et al, 2019). LPS causes a significant upregulation of TNF-α and IL-1β (Chen et al, 2013;Ni et al, 2019), upregulating MMPs gene expression and proinflammatory cytokine, driving apoptosis via the effector caspases, causing cartilage degradation (Kaneva et al, 2012(Kaneva et al, , 2014.…”

The nuclear factor-erythroid 2-related factor/heme oxygenase-1 axis is critical for the inflammatory features of type 2 diabetes–associated osteoarthritis

“…However, in response to oxidative or electrophilic stress, Nrf-2 dissociates from Keap1 and translocates to the nucleus to bind antioxidant responsive elements (ARE) in the promoter regions of its downstream antioxidant genes, including heme oxygenase-1 (HO-1) (24). HO-1 is a crucial antioxidant enzyme that catalyzes the degradation of heme into iron, carbon monoxide, and biliverdin (25). This enzyme regulates catabolic and anabolic processes in OA chondrocytes (26).…”

Carnosic acid attenuates cartilage degeneration through induction of heme oxygenase-1 in human articular chondrocytes

“…These findings suggest that maintaining, HO-1 expression may have beneficial effects for OA prevention during aging. Overexpression or induction of HO-1 has anti-inflammatory activities (Benallaoua et al, 2007; Mawatari et al, 2013; Ryter and Choi, 2016). The induction of HO-1 by CoPP protects against inflammatory and degradative responses in OA chondrocytes and synoviocytes (Clérigues et al, 2013; García-Arnandis et al, 2010; Guillén et al, 2008; Megías et al, 2009).…”