Immortalized human adult articular chondrocytes maintain cartilage-specific phenotype and responses to interleukin-1β

Robbins, J B; Thomas, Beatrice C.; Tan, Lujian; Choy, Bob K.; Arbiser, Jack L.; Bérenbaum, Francis; Goldring, Mary B.

doi:10.1002/1529-0131(200010)43:10<2189::aid-anr6>3.0.co;2-s

Cited by 107 publications

(77 citation statements)

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“…Since only limited amounts of human tissue were available, we used the immortalized chondrocyte cell line T/C-28a2, which retains the characteristics of chondrocytes and provides enough cells for extensive studies (24,25,35). We used Northern blotting Figure 2B).…”

Section: Resultsmentioning

confidence: 99%

Up‐regulation of microsomal prostaglandin E synthase 1 in osteoarthritic human cartilage: Critical roles of the ERK‐1/2 and p38 signaling pathways

Masuko

Bérenbaum

Humbert

et al. 2004

Arthritis & Rheumatism

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118

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Objective. Microsomal prostaglandin E synthase 1 (mPGES-1) is the final enzyme of the cascade that produces prostaglandin E 2 (PGE 2 ), a key actor in arthritis. To study mPGES-1 synthesis in human cartilage and its regulation by interleukin-1␤ (IL-1␤), we used human cartilage and an immortalized human chondrocyte cell line. Furthermore, we investigated the signaling pathways involved in mPGES-1 expression.Methods. We used real-time quantitative reverse transcription-polymerase chain reaction, Northern blotting, and Western blotting to measure mPGES-1 messenger RNA (mRNA) and protein expression in human chondrocytes. PGE 2 production was measured by enzyme-linked immunosorbent assay.Results. Cartilage specimens from osteoarthritis (OA) patients contained far greater amounts of mPGES-1 and cyclooxygenase 2 (COX-2) mRNA than did normal cartilage. Incubation with IL-1␤ markedly increased mPGES-1 mRNA and protein in a dosedependent and time-dependent manner, in parallel with an increase in PGE 2 levels. Both PD98059, an ERK pathway inhibitor, and SB203580, a p38␣/␤ MAPK inhibitor, abolished the increases in mPGES-1 mRNA and protein in response to IL-1␤. The specific p38␣ MAPK inhibitor SC906 suppressed IL-1␤-induced COX-2 expression but not IL-1␤-induced mPGES-1 expression, suggesting preferential involvement of p38␤ MAPK in IL-1␤-induced mPGES-1 expression.Conclusion. This study is the first to show that mPGES-1 is stimulated in human chondrocytes by the proinflammatory cytokine IL-1␤ via activation of both ERK-1/2 and p38 MAPK in an isoform-specific manner. We postulate that mPGES-1 may be a novel target for OA therapy.Prostaglandin E 2 (PGE 2 ) plays an important role in cartilage metabolism. Its many effects on chondrocytes (for review, see ref. 1) include enhanced production of matrix metalloproteinase 3 (2), modulation of proteoglycan and collagen synthesis (2,3), and stimulation of chondrocyte apoptosis (4,5). The synovial fluid of patients with osteoarthritis (OA) and rheumatoid arthritis contains high concentrations of PGE 2 (4), and cartilage and chondrocytes from OA patients spontaneously release more PGE 2 than does normal cartilage (2,6,7). These results suggest that PGE 2 may be actively involved in cartilage breakdown in OA patients. PGE 2 is synthesized by the isoenzymes cyclooxygenase 1 (COX-1) and COX-2, both of which act on arachidonic acid to form the prostaglandin endoperoxide H 2 (PGH 2 ). The prostanoid synthase prostaglandin E synthase (PGES) produces PGE 2 from PGH 2 . Three

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Section: Resultsmentioning

confidence: 99%

Up‐regulation of microsomal prostaglandin E synthase 1 in osteoarthritic human cartilage: Critical roles of the ERK‐1/2 and p38 signaling pathways

Masuko

Bérenbaum

Humbert

et al. 2004

Arthritis & Rheumatism

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118

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“…More recently, IL-1␤ was even demonstrated to support cartilage damage in human TNF-␣-transgenic mice possibly due to its regulatory role on matrix-degrading enzymes (32). At the cellular level, IL-1␤ was shown to be a potent modulator of the chondrocyte phenotype, as it down-regulated the expression of some cartilage-specific genes including those encoding Types IX (33), XI, and II (34) collagens and aggrecan (34). Most of FIGURE 3.…”

Section: Discussionmentioning

confidence: 99%

The Inorganic Pyrophosphate Transporter ANK Preserves the Differentiated Phenotype of Articular Chondrocyte

Cailotto

Sébillaud

Netter

et al. 2010

Journal of Biological Chemistry

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The differentiated phenotype of chondrocyte is lost in pathological situations and after interleukin (IL)-1␤ challenge. Wnt proteins and the inorganic pyrophosphate (PP i ) transporter Ank regulate the differentiation process in many cell types. We investigated the possible contribution of Ank and/or PP i to the maintenance of the differentiated chondrocyte phenotype with special care to Wnt signaling. Primary articular chondrocytes lost their phenotype upon IL-1␤ challenge, with cessation of type II collagen and Sox-9 expression. Ank expression and PP i transport were strongly reduced by IL-1␤, whereas Wnt-5a was the only Wnt protein increased. Transient overexpression of Ank counteracted most of IL-1␤ effects on Type II collagen, Sox-9, and Wnt-5a expression. When resting chondrocytes were transfected with a siRNA against Ank, this reproduced the phenotype induced by IL-1␤. In both cases, no markers for hypertrophic chondrocytes were detected. The conditioned supernatant from chondrocytes knocked-down for Ank contained Wnt-5a, which activated Tcf/Lef reporter plasmids and promoted translocation of ␤-catenin into the nucleus without activating the c-Jun N-terminal kinase (JNK) pathway. Supplementation with PP i compensated for most effects of Ank deficiency on Type II collagen, Sox-9, and Wnt-5 expression, both in IL-1␤ and Ank knock-down conditions. Phenotype changes induced by IL-1␤ were also supported by activation of the JNK pathway, but this latter was not sensitive to PP i supplementation. Altogether our data demonstrate that the transport of PP i by ANK contributed to the maintenance of the differentiated phenotype of chondrocyte by controlling the canonical Wnt pathway in a Wnt-5a-dependent manner.

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“…Several lines of evidence suggest that cytokines, such as IL-1b and TNF-a, are produced in high quantities in OA and RA in cartilage. Once released, they stimulate the synthesis of more pro-inflammatory cytokines, catabolic factors such as matrix metalloproteinases (MMPs) [22,39,40] and mediators of inflammation such as prostaglandins produced by cyclooxygenase-2 (COX-2) [28]. Pro-inflammatory cytokines also induce chondrocyte apoptosis [11], a process that is thought to play a pivotal role in joint diseases (OA and RA) in humans and animals [3,24,25].…”

Section: Introductionmentioning

confidence: 99%

Curcumin synergizes with resveratrol to stimulate the MAPK signaling pathway in human articular chondrocytes in vitro

2010

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The mitogen-activated protein kinase (MAPK) pathway is stimulated in differentiated chondrocytes and is an important signaling cascade for chondrocyte differentiation and survival. Pro-inflammatory cytokines such as interleukin 1b (IL-1b) play important roles in the pathogenesis of osteoarthritis (OA) and rheumatoid arthritis (RA). In this study, we investigated whether curcumin and resveratrol can synergistically inhibit the catabolic effects of IL-1b, specifically the inhibition of the MAPK and subsequent apoptosis in human articular chondrocytes. Chondrocytes were either left untreated or treated with 10 ng/ml IL-1b or 1 lM U0126, a specific inhibitor of MAPK pathway alone for the indicated time periods or pretreated with 10 lM curcumin, 10 lM resveratrol or 10 lM resveratrol and 10 lM curcumin for 4 h followed by cotreatment with 10 ng/ml IL-1b or 1 lM U0126 and 10 lM resveratrol, 10 lM curcumin or 10 lM resveratrol and 10 lM curcumin for the indicated time periods. Cultures were evaluated by immunoblotting and transmission electron microscopy. Incubation of chondrocytes with IL-1b resulted in induction of apoptosis, downregulation of b1-integrins and the extracellular signal-regulated kinase 1/2 (Erk1/2). Interestingly, U0126 induced apoptosis and blocked the above-mentioned proteins in a similar way to IL-1b. Furthermore, curcumin and resveratrol inhibited IL-1b-or U0126-induced apoptosis and downregulation of b1-integrins and Erk1/2 in human articular chondrocytes. These results suggest that combining these two natural compounds activates MEK/Erk signaling, a pathway that is involved in the maintenance of chondrocyte differentiation and survival.

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Immortalized human adult articular chondrocytes maintain cartilage-specific phenotype and responses to interleukin-1β

Cited by 107 publications

References 50 publications

Up‐regulation of microsomal prostaglandin E synthase 1 in osteoarthritic human cartilage: Critical roles of the ERK‐1/2 and p38 signaling pathways

Up‐regulation of microsomal prostaglandin E synthase 1 in osteoarthritic human cartilage: Critical roles of the ERK‐1/2 and p38 signaling pathways

The Inorganic Pyrophosphate Transporter ANK Preserves the Differentiated Phenotype of Articular Chondrocyte

Curcumin synergizes with resveratrol to stimulate the MAPK signaling pathway in human articular chondrocytes in vitro

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