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Cited by 182Osteoarthritis Treatment and MechanismsProteoglycans and glycosaminoglycans researchProtease and Inhibitor Mechanisms

Arthropod steroid hormone (20-Hydroxyecdysone) suppresses IL-1β- induced catabolic gene expression in cartilage

Open access
Sheu, Shiow-Yunn, Ho, Shin-Rong, Sun, Jui-Sheng, Chen, Ching-Yun, Ke, Cherng-Jyh·2015·BMC Complementary and Alternative Medicine
DOI 10.1186/s12906-015-0520-zPMC PMC4310028
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Abstract

BACKGROUND: In osteoarthritis (OA), the imbalance of chondrocytes' anabolic and catabolic factors can induce cartilage destruction. Interleukin-1 beta (IL-1β) is a potent pro-inflammatory cytokine that is capable of inducing chondrocytes and synovial cells to synthesize MMPs. The hypoxia-inducible factor-2alpha (HIF-2alpha, encoded by Epas1) is the catabolic transcription factor in the osteoarthritic process. The purpose of this study is to validate the effects of ecdysteroids (Ecd) on IL-1β-induced cartilage catabolism and the possible role of Ecd in treatment or prevention of early OA. METHODS: Chondrocytes and articular cartilage was harvested from newborn ICR mice. Ecd effect on chondrocytes viability was tested and the optimal concentration was determined by MTT assay. The effect of HIF-2α (EPAS1) in cartilage catabolism simulated by IL-1β (5 ng/ml) was evaluated by articular cartilage explants culture. The effects of Ecd on IL-1β-induced inflammatory conditions and their related catabolic genes expression were analyzed. RESULTS: Interleukin-1β (IL-1β) treatment on primary mouse articular cartilage explants enhanced their Epas1, matrix metalloproteinases (MMP-3, MMP-13) and ADAMTS-5 genes expression and down-regulated collagen type II (Col2a1) gene expression. With the pre-treatment of 10(-8) M Ecd, the catabolic effects of IL-1β on articular cartilage were scavenged. CONCLUSION: In conclusions, Ecd can reduce the IL-1β-induced inflammatory effect of the cartilage. Ecd may suppress IL-1β-induced cartilage catabolism via HIF-2α pathway.

AI snapshot

Written from the paper’s full text.

TL;DR

In newborn ICR mouse chondrocytes and articular cartilage explants, IL-1β (5 ng/ml) increased Epas1, MMP-3, MMP-13, and ADAMTS-5 and decreased Col2a1; pretreatment with 10−8 M 20-hydroxyecdysone suppressed these catabolic changes. The authors conclude that Ecd has chondroprotective effects via the HIF-2α pathway, but call for further in vivo and translational work.

Key findings

  • Ecd enhanced chondrocyte viability, with maximal benefit at 10−8 M after 10 days of culture (p < 0.01); significant differences from control were observed at tested Ecd concentrations (p < 0.05), and 10−8 M was chosen for further experiments.
  • IL-1β treatment enhanced Epas1 mRNA and suppressed Col2a1 in chondrocytes; in articular cartilage explants, IL-1β up-regulated MMP-3, MMP-13, and ADAMTS-5 and down-regulated Col2a1. Pretreatment with 10−8 M Ecd eliminated the IL-1β-enhanced Epas1 expression and down-regulated the catabolic genes, but did not affect Col2a1 in the conditions shown in Figure 2.
  • Under hypoxic conditions (3.5% O2), Epas1 expression was higher; Ecd suppressed IL-1β-induced Epas1 and ADAMTS-5 under both hypoxic and normoxic conditions, while MMP-3 was down-regulated only under normoxic conditions. Under hypoxia, IL-1β-mediated ADAMTS-5 up-regulation was the main catabolic mediator, and no significant MMP13 expression was found (data not shown).

Methods

  • Primary chondrocytes and articular cartilage explants were harvested from newborn ICR mice (3 days old); chondrocytes were isolated by collagenase digestion and cultured. Viability was tested by MTT assay with Ecd at 10−6 to 10−10 M on days 1, 3, 7, and 10 (n = 8).
  • Cartilage explants were pre-incubated with or without Ecd (10−8 M), then treated with IL-1β (5 ng/ml); Ecd was added 24 hours before or after IL-1β, and explants were incubated under hypoxic (3.5% O2) or normoxic conditions.
  • Gene expression was analyzed by RT-PCR for Col2a1, MMP-3, MMP-13, ADAMTS5, EPAS1, and β-actin at 0, 3, 6, 12, 24, and 36 hours after Ecd addition. Experiments were performed at least in triplicate; results were analyzed by two-way ANOVA with Dunnett’s test at p < 0.05.

Limitations

  • The authors state that further in vivo HIF-1alpha gene knock-down and translational medicine studies are mandatory for possible clinical application.
  • Evident from the text: the study is limited to in vitro mouse chondrocyte cultures and articular cartilage explants, with no in vivo osteoarthritis model, protein/activity assays, or histological outcomes reported.
  • Evident from the text: Ecd did not affect Col2a1 in Figure 2, and MMP13 expression under hypoxic conditions was not shown, limiting conclusions about full phenotypic recovery and hypoxic MMP13 involvement.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1186/s12906-015-0520-z

Cites

34 of 34 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 28 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Under hypoxic condition, IL-1β up-regulates ADAMTS-5 gene expression in articular cartilage.

    “Under hypoxic condition, IL-1β mediates HIF-2α–induced up-regulation of ADAMTS-5 gene (but not MMP3 or MMP13 genes) in articular cartilage (Figure 4C and D)”
    Quote in the paperSubstantial claim
  • Findingown data

    Under hypoxic condition, IL-1β does not significantly up-regulate MMP-13 gene expression in articular cartilage.

    “there was no significant expression on MMP13 gene under hypoxic condition (data not shown).”
    Quote in the paperSubstantial claim
  • Findingown data

    Pre-treatment with 10^-8 M Ecd down-regulates IL-1β-induced ADAMTS-5 gene expression in articular cartilage explants.

    “with the pretreatment of 10−8M Ecd, the catabolic factor genes upregulated by IL-1β treatment are eliminated.”
    Quote in the paperSubstantial claim
  • Findingown data

    Under hypoxic condition, IL-1β does not significantly up-regulate MMP-3 gene expression in articular cartilage.

    “Under hypoxic condition, IL-1β mediates HIF-2α–induced up-regulation of ADAMTS-5 gene (but not MMP3 or MMP13 genes) in articular cartilage (Figure 4C and D)”
    Quote in the paperSubstantial claim
  • Findingown data

    IL-1β treatment enhances Epas1 gene expression in primary mouse articular cartilage explants.

    “Same as noted above, IL-1β enhances Epas1 mRNA expression and suppresses collagen type II (Col2a1) gene expression.”
    Quote in the paperSubstantial claim
  • The quote only partly states this claim. 1
  • May combine two claims. 5
  • Classified as probably not a substantial claim. 13

  • The quote only partly states this claim.

    Findingown data

    Ecd pre-treatment does not affect Col2a1 gene expression in articular cartilage explants under normoxic conditions.

    “but there is also no effect on Col2a1 gene expression (Figure 2).”
    • Also stated in: § Discussion · plain
      “but there is no ability to recover the cartilage phenotype as Col2a1 gene was not affected (Figure 4).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    In ovariectomized rats, pure Ecd supplementation increases joint cartilage thickness and has beneficial bone effects.

    “In ovariectomized rats, pure Ecd supplementation significantly increased in the thickness of joint cartilage and presented beneficial effects in bones.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [13] Kapur P, Wuttke W, Jarry H, Seidlova-Wuttke D. Beneficial effects of beta-Ecdysone on the joint, epiphyseal cartilage tissue and trabecular bone in ovariectomized rats. Phytomedicine. 2010;17(5):350–5. source

      Not checked in the free run.

    • cites [14] Zhang X, Xu X, Xu T, Qin S. β-Ecdysterone suppresses interleukin-1β-induced apoptosis and inflammation in rat chondrocytes via inhibition of NF-κB signaling pathway. Drug Dev Res. 2014;75(3):195–201. source

      Not checked in the free run.

    • cites [15] He CC, Hui RR, Tezuka Y, Kadota S, Li JX. Osteoprotective effect of extract from Achyranthes bidentata in ovariectomized rats. J Ethnopharmacol. 2010;127(2):229–34. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    IL-1β and other inflammatory mediators promote activation of latent ADAMTS-5.

    “Instead, IL-1β and certain other inflammatory mediators promote activation of latent ADAMTS-5, as does hypertrophic differentiation in chondrocytes [3,4].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [3] Saito T, Fukai A, Mabuchi A, Ikeda T, Yano F, Ohba S, et al. Transcriptional regulation of endochondral ossification by HIF-2alpha during skeletal growth and osteoarthritis development. Nat Med. 2010;16(6):678–86. source

      Not checked in the free run.

    • cites [4] Yang S, Kim J, Ryu JH, Oh H, Chun CH, Kim BJ, et al. Hypoxia-inducible factor-2alpha is a catabolic regulator of osteoarthritic cartilage destruction. Nat Med. 2010;16(6):687–93. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    MMP-3 is strongly expressed in normal and early degenerative cartilage and down-regulated in late disease stages.

    “In spite of a large variability of expression levels among different patients, MMP-3 appeared to be strongly expressed in normal and early degenerative cartilage and down-regulated in the late disease stages.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] Aigner T, Zien A, Hanisch D, Zimmer R. Gene expression in chondrocytes assessed with use of microarrays. J Bone Joint Surg Am. 2003;85-A Suppl 2:117–23. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Ecd has no known adverse effects even when taken in gram quantities.

    “The Ecd is frequently used by body builders and from these users it is known that Ecd has no known adverse effects even when taken up in gram (~1 g) quantities [16].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Dinan L, Lafont R. Effects and applications of arthropod steroid hormones (ecdysteroids) in mammals. J Endocrinol. 2006;191(1):1–8. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    HIF-2α does not induce transcription of ADAMTS5.

    “It is reported that HIF-2α (like IL-1β) does not induce transcription of ADAMTS5 [3,4].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [3] Saito T, Fukai A, Mabuchi A, Ikeda T, Yano F, Ohba S, et al. Transcriptional regulation of endochondral ossification by HIF-2alpha during skeletal growth and osteoarthritis development. Nat Med. 2010;16(6):678–86. source

      Not checked in the free run.

    • cites [4] Yang S, Kim J, Ryu JH, Oh H, Chun CH, Kim BJ, et al. Hypoxia-inducible factor-2alpha is a catabolic regulator of osteoarthritic cartilage destruction. Nat Med. 2010;16(6):687–93. source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    A stress-induced increase in HIF-2α activity overshadows HIF-1α and drives osteoarthritis changes.

    “According to this paradigm, a stress-induced increase in the activity of HIF-2α will overshadow the beneficial effects of the closely related HIF-1α, then push the chondrocytes in the joint toward a more differentiated state known as hypertrophy, which then drives osteoarthritis changes [6,7].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [6] Husa M, Liu-Bryan R, Terkeltaub R. Shifting HIFs in osteoarthritis. Nat Med. 2010;16(6):641–4. source

      Not checked in the free run.

    • cites [7] Ryu JH, Shin Y, Huh YH, Yang S, Chun CH, Chun JS. Hypoxia-inducible factor-2α regulates Fas-mediated chondrocyte apoptosis during osteoarthritic cartilage destruction. Cell Death Differ. 2012;19(3):440–50. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Hypoxic signaling is a central regulator of skeletal biology and disease.

    “Together, these findings implicate hypoxic signaling as a central regulator of skeletal biology and disease [32].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [32] Rankin EB, Giaccia AJ, Schipani E. A central role for hypoxic signaling in cartilage, bone, and hematopoiesis. Curr Osteoporos Rep. 2011;9(2):46–52. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    Inflammation of articular cartilage stimulates ADAMTS5 activation.

    “The findings of our study are consistent with the observations that inflammation of articular cartilage stimulates ADAMTS5 activation.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    ADAMTS-4 and ADAMTS-5 are implicated prominently in cartilage breakdown during OA.

    “During OA, ADAMTS-4 (aggrecanase 1) [28] and ADAMTS-5 (aggrecanase 2) [29,30] have been implicated prominently in cartilage breakdown.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [28] Poole AR, Nelson F, Dahlberg L, Tchetina E, Kobayashi M, Yasuda T, et al. Proteolysis of the collagen fibril in osteoarthritis. Biochem Soc Symp. 2003;70:115–23. source

      Not checked in the free run.

    • cites [29] Tortorella MD, Burn TC, Pratta MA, Abbaszade I, Hollis JM, Liu R, et al. Purification and cloning of aggrecanase-1: a member of the ADAMTS family of proteins. Science. 1999;284(5420):1664–6. source

      Not checked in the free run.

    • cites [30] Pattoli MA, MacMaster JF, Gregor KR, Burke JR. Collagen and aggrecan degradation is blocked in interleukin-1-treated cartilage explants by an inhibitor of IkappaB kinase through suppression of metalloproteinase expression. J Pharmacol Exp Ther. 2005;315(1):382–8. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Degradation of aggrecan by aggrecanases is an early event in osteoarthritic cartilage damage followed by irreversible loss of collagens.

    “The degradation of aggrecan by aggrecanases has gained special interest, because it constitutes an early event in osteoarthritic cartilage damage and is followed by the irreversible loss of collagens [27].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [27] Brooks PM. The burden of musculoskeletal disease–a global perspective. Clin Rheumatol. 2006;25(6):778–81. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    For each gene analysis, experiments were repeated at least three times.

    “For each genes analysis, the experiments were repeated at least three times.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Ecd has protein anabolic effects and increases muscle mass in mammals.

    “In mammals, Ecd has protein anabolic effects and increases muscle mass of unexercised and exercised muscle [11,12].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [11] Báthori M, Tóth N, Hunyadi A, Márki A, Zádor E. Phytoecdysteroids and anabolic-androgenic steroids–structure and effects on humans. Curr Med Chem. 2008;15(1):75–91. source

      Not checked in the free run.

    • cites [12] Gorelick-Feldman J, Maclean D, Ilic N, Poulev A, Lila MA, Cheng D, et al. Phytoecdysteroids increase protein synthesis in skeletal muscle cells. J Agric Food Chem. 2008;56(10):3532–7. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    All experiments were performed at least in triplicate.

    “All experiments were performed at least in triplicate.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    MMP-13 is predominantly expressed in lower intermediate and deep layers of cartilage in early OA, while aggrecanases are expressed in superficial cartilage.

    “In the early phases of OA, MMP-13 is predominantly expressed in the lower intermediate and deep layers of human and mouse cartilage, whereas aggrecanases are expressed in the superficial cartilage [25,26].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [25] Brew CJ, Clegg PD, Boot-Handford RP, Andrew JG, Hardingham T. Gene expression in human chondrocytes in late osteoarthritis is changed in both fibrillated and intact cartilage without evidence of generalised chondrocyte hypertrophy. Ann Rheum Dis. 2010;69(1):234–40. source

      Not checked in the free run.

    • cites [26] Little CB, Barai A, Burkhardt D, Smith SM, Fosang AJ, Werb Z, et al. Matrix metalloproteinase 13-deficient mice are resistant to osteoarthritic cartilage erosion but not chondrocyte hypertrophy or osteophyte development. Arthritis Rheum. 2009;60(12):3723–33. source

      Not checked in the free run.

15 of 15

  • The paper relies on this without support.

    Epas1 gene expression is a valid proxy for HIF-2α activity.

    “The hypoxia-inducible factor-2alpha (HIF-2α, encoded by Epas1) is a catabolic transcription factor in the osteoarthritic process.”

  • The paper relies on this without support.

    Findings in mouse articular cartilage explants are relevant to human OA.

    “The purpose of this study is to validate the effects of Ecd on IL-1β- induced cartilage destruction and also the possible role of Ecd in treatment or prevention of early OA.”
2 of 2

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • 20E scavenged the increase in catabolic processes and decrease in anabolic processes and largely restored the balance in primary explanted articular chondrocytes.

    “In addition, 20E scavenged the increase in catabolic processes and decrease in anabolic processes and largely restored the balance in primary explanted articular chondrocytes [39], confirming a protective effect in OA.”

    From Molecular Targets of 20-Hydroxyecdysone in Mammals, Mechanism of Action: Is It a Calorie Restriction Mimetic and Anti-Aging Compound? (2025) · cites this paper as [39]

    The Results section reports that 20E pretreatment reduced IL-1β-induced catabolic gene expression in primary mouse articular cartilage explants. It also says 20E had no effect on collagen type II (Col2a1) gene expression, which IL-1β had suppressed. Thus the source supports a protective effect against the measured catabolic changes, but not restoration of the decreased anabolic process or the claimed balance. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

Cites

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Related

Authors
Sheu, S.-Y., Ho, S.-R., Sun, J.-S., Chen, C.-Y., & Ke, C.-J.
Affiliations
  • Taipei Medical University (TW)
  • National Yang Ming Chiao Tung University (TW)
  • National Taiwan University (TW)
Corresponding author
Sun, Jui-Sheng
ORCID
Venue
BMC Complementary and Alternative Medicine
Publisher
Springer Science and Business Media LLC
Published
2015-01-24
Volume
15
Issue
1
Pages
1-1
ISSN
1472-6882 (ISSN-L)
DOI
10.1186/s12906-015-0520-z
PMC
PMC4310028
OpenAlex
W2005726309
Semantic Scholar
2d21a907221820c599a0466437d4df9f7688707f
Cited by
181
References
34
Citations by year
2026: 4 · 2025: 18 · 2024: 15 · 2023: 14 · 2022: 11 · 2021: 29 · 2020: 16 · 2019: 19 · 2018: 17 · 2017: 19 · 2016: 17 · 2015: 2
FWCI
15.36
Type
journal article
Language
en
Open access
Licence
Locations
  • BMC Complementary and Alternative Medicine (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Science Council
  • National Taiwan University Hospital
  • National Taiwan University
Topics
  • Health Sciences › Medicine › Rheumatology › Osteoarthritis Treatment and Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Proteoglycans and glycosaminoglycans research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Protease and Inhibitor Mechanisms
Keywords
20-hydroxyecdysone, ecdysteroids, osteoarthritis, cartilage catabolism, IL-1β, HIF-2α, EPAS1, chondrocytes, matrix metalloproteinases, ADAMTS5, type II collagen, articular cartilage, inflammation
MeSH terms
Endothelial PAS Domain-Containing Protein 1, Animals, Arthropods, Cartilage, Cartilage, Articular, Cells, Cultured, Ecdysterone, Male, Mice, Inbred ICR, Osteoarthritis, Synovial Membrane, Transcription Factors, Down-Regulation, Gene Expression, Cytokines, Matrix Metalloproteinase 3, Chondrocytes, Collagen Type II, Basic Helix-Loop-Helix Proteins, Matrix Metalloproteinase 13, Interleukin-1beta

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