Modulation of cytokine-induced prostaglandin E2 production in cultures of articular chondrocytes obtained from carpal joints of camels (Camelus dromedarius)

Frondoza, Carmelita G.; Heinecke, L.F.; Grzanna, M.W.; Au, A.Y.; Ownby, Stacy

doi:10.2460/ajvr.72.1.51

Cited by 8 publications

(12 citation statements)

References 41 publications

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“…ASU and EGCG have both been documented for their anti-inflammatory activity [7,8,11,12]. Of the two compounds, ASU has been reported to have pro-anabolic activity and has been used for OA management in both humans and animals [6,24].…”

Section: Resultsmentioning

confidence: 99%

“…Following pre-treatment, cultures were incubated with control media alone or activated with human recombinant cytokines IL-1β (10 ng/mL; R&D Systems, Minneapolis, MN) and TNF-α (1 ng/mL; Sigma-Aldrich) for 1 h to measure gene expression by real-time PCR, NF-κB immunohistochemistry, or Western blot. Earlier studies have confirmed the combination of IL-1β + TNF-α at these concentrations to be a potent activator for chondrocytes from several different species including equine and camel [8,16]. Following activation, supernatant was removed and a set of 6-well plates was stored at -80°C for gene expression analysis while another set was used for nuclear fractionation.…”

Section: Methodsmentioning

confidence: 99%

“…Nuclear translocation of NF-κB was monitored by immunohistochemistry as previously described using a primary antibody specific to the p65 subunit of NF-κB (rabbit anti-NF-κB; Santa Cruz Biotechnology, Santa Cruz, CA), which we earlier determined to cross-react with equine and camel chondrocytes [8,16]. System controls were included which consisted of normal serum to substitute for the primary specific antibody in which only background staining was observed.…”

Section: Methodsmentioning

confidence: 99%

“…ASU suppresses gene expression of IL-1β, TNF-α, COX-2, and inducible nitric oxide synthase (iNOS), as well as PGE 2 and nitric oxide production in bovine and human joint tissue cells [6,7]. When combined with other compounds such as glucosamine, chondroitin sulfate, and pentosan polysulfate, ASU potentiated anti-inflammatory activity [8,9]. Another compound reported to have anti-inflammatory activity is epigallocatechin gallate (EGCG).…”

Section: Introductionmentioning

confidence: 99%

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Expression of pro-inflammatory mediators is inhibited by an avocado/soybean unsaponifiables and epigallocatechin gallate combination

Ownby

Fortuno

et al. 2014

Journal of Inflammation

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BackgroundOsteoarthritis (OA) is characterized by inflammation, joint immobility, and pain. Non-pharmacologic agents modulating pro-inflammatory mediator expression offer considerable promise as safe and effective treatments for OA. We previously determined the anti-inflammatory effect of an avocado/soybean unsaponifiables (ASU) and epigallocatechin gallate (EGCG) combination on prostaglandin E2 (PGE2) production and nuclear factor-kappa B (NF-κB) translocation. The aim of this study was to evaluate the effects of ASU + EGCG on pro-inflammatory gene expression.FindingsArticular chondrocytes from carpal joints of mature horses were pre-incubated for 24 hours with control media alone or ASU (8.3 μg/mL) + EGCG (40 ng/mL), followed by one hour activation with interleukin-1 beta (IL-1β, 10 ng/mL) and tumor necrosis factor-alpha (TNF-α, 1 ng/mL). Total cellular RNA was isolated and real-time PCR performed to measure IL-1β, TNF-α, interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), and interleukin-8 (IL-8) gene expression. Intracellular localization of NF-κB was analyzed by immunohistochemistry and Western blot. Pre-treatment with ASU + EGCG significantly (P < 0.001) decreased gene expression of IL-1β, TNF-α, IL-6, COX-2, and IL-8 in cytokine-activated chondrocytes. Western blot and immunostaining confirmed NF-κB translocation inhibition.ConclusionsWe demonstrate that ASU + EGCG inhibits cytokine-induced gene expression of IL-1β, TNF-α, IL-6, COX-2, and IL-8 through modulation of NF-κB. Our results indicate that the activity of ASU + EGCG affects a wide array of inflammatory molecules in addition to decreasing PGE2 synthesis in activated chondrocytes. The responsiveness of chondrocytes to this combination supports its potential utility for the inhibition of joint inflammation.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Expression of pro-inflammatory mediators is inhibited by an avocado/soybean unsaponifiables and epigallocatechin gallate combination

Ownby

Fortuno

et al. 2014

Journal of Inflammation

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“…ASU, GLU and CS have been documented to reduce inflammation in vitro [9] [10] [11], [31]- [38], and in vivo [39]- [46]. These agents have been reported to ameliorate OA in man and animals with minimal adverse side effects [39]- [46].…”

Section: Introductionmentioning

confidence: 99%

Chondrocyte Production of Pro-Inflammatory Chemokine MCP-1 (CCL-2) and Prostaglandin E-2 Is Inhibited by Avocado/Soybean Unsaponifiables, Glucosamine, Chondroitin Sulfate Combination

Secor¹,

Grzanna²,

Rashmir‐Raven³

et al. 2018

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Osteoarthritis (OA) is a chronic, painful disease affecting articulating joints in man and animals. It is characterized by cartilage breakdown, bone remodeling, osteophyte formation and joint inflammation. Currently used non-steroidal anti-inflammatory drugs for the management of OA are known to have deleterious side effects. To address the need for alternatives, we evaluated the anti-inflammatory effects of a combination of avocado/soybean unsaponifiables (ASU), glucosamine (GLU) and chondroitin sulfate (CS) by measuring chemokine MCP-1 (monocyte chemoattractant protein 1, CCL2) and prostaglandin E-2 (PGE 2 ) in stimulated chondrocytes. As the only cellular constituents of cartilage, chondrocytes are the source of pro-inflammatory mediators that play critical roles in the pathogenesis of OA. Chondrocytes were incubated: with: 1) control media, 2) [ASU + GLU + CS] combination, or 3) Phenylbutazone (PBZ) for 24 hours. Cells were next stimulated with IL-1β or LPS for another 24 hrs. MCP-1 and PGE 2 from supernatants were quantitated by immunoassay. Another set of chondrocytes seeded in chamber slides were stimulated with IL-1β for 1 hour and then immunostained for NF-κB. Chondrocytes stimulated with IL-1β or LPS significantly increased MCP-1 and PGE 2 production which were significantly reduced after treatment with [ASU + GLU + CS]. In contrast, PBZ significantly reduced PGE 2 but not MCP-1 production. IL-1β stimulation induced nuclear translocation of NF-κB, which The present study provides evidence that the production of MCP-1 by chondrocytes can be inhibited by the combination of [ASU + GLU + CS] but not by PBZ. In contrast, PGE 2 production was inhibited by either treatment suggesting that the production of MCP-1 and PGE 2 could be independently regulated. The finding of distinct effects of [ASU + GLU + CS] on MCP-1 and PGE 2 synthesis supports a scientific rationale for a multimodal treatment approach in the management of OA.

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Avocado/Soybean Unsaponifiables, Glucosamine and Chondroitin Sulfate Combination Inhibits Proinflammatory COX-2 Expression and Prostaglandin E2 Production in Tendon-Derived Cells

Grzanna

et al. 2020

Journal of Medicinal Food

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Modulation of cytokine-induced prostaglandin E2 production in cultures of articular chondrocytes obtained from carpal joints of camels (Camelus dromedarius)

Cited by 8 publications

References 41 publications

Expression of pro-inflammatory mediators is inhibited by an avocado/soybean unsaponifiables and epigallocatechin gallate combination

Expression of pro-inflammatory mediators is inhibited by an avocado/soybean unsaponifiables and epigallocatechin gallate combination

Chondrocyte Production of Pro-Inflammatory Chemokine MCP-1 (CCL-2) and Prostaglandin E-2 Is Inhibited by Avocado/Soybean Unsaponifiables, Glucosamine, Chondroitin Sulfate Combination

Avocado/Soybean Unsaponifiables, Glucosamine and Chondroitin Sulfate Combination Inhibits Proinflammatory COX-2 Expression and Prostaglandin E2 Production in Tendon-Derived Cells

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