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Cited by 30Bone health and osteoporosis researchBone Metabolism and DiseasesMedicinal Plant Pharmacodynamics Research

Prevention of glucocorticoid induced bone changes with beta-ecdysone

Open access
Dai, Weiwei, Jiang, Li, Lay, Yu-An Evan, Chen, Haiyan, Jin, Guoqin, Zhang, Hongliang, Kot, Alexander, Ritchie, Robert O. +2 more·2015·Bone
DOI 10.1016/j.bone.2015.01.001PMC PMC4355031
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Abstract

Beta-ecdysone (βEcd) is a phytoecdysteroid found in the dry roots and seeds of the asteraceae and achyranthes plants, and is reported to increase osteogenesis in vitro. Since glucocorticoid (GC) excess is associated with a decrease in bone formation, the purpose of this study was to determine if treatment with βEcd could prevent GC-induced osteoporosis. Two-month-old male Swiss-Webster mice (n=8-10/group) were randomized to either placebo or slow release prednisolone pellets (3.3mg/kg/day) and treated with vehicle control or βEcd (0.5mg/kg/day) for 21days. GC treatment inhibited age-dependent trabecular gain and cortical bone expansion and this was accompanied by a 30-50% lower bone formation rate (BFR) at both the endosteal and periosteal surfaces. Mice treated with only βEcd significantly increased bone formation on the endosteal and periosteal bone surfaces, and increased cortical bone mass were their controls to compare to GC alone. Concurrent treatment of βEcd and GC completely prevented the GC-induced reduction in BFR, trabecular bone volume and partially prevented cortical bone loss. In vitro studies determined that βEcd prevented the GC increase in autophagy of the bone marrow stromal cells as well as in whole bone. In summary, βEcd prevented GC induced changes in bone formation, bone cell viability and bone mass. Additional studies are warranted of βEcd for the treatment of GC induced bone loss.

AI snapshot

Written from the paper’s full text.

TL;DR

In 2-month-old male Swiss-Webster mice, β-ecdysone (βEcd) increased bone formation and bone mass and, when co-administered with prednisolone, prevented glucocorticoid (GC)-induced reductions in trabecular bone volume and bone formation rate while partially preventing cortical bone loss. In vitro and in vivo assays suggest βEcd may act partly by sustaining osteoblastic autophagy that GC suppresses.

Key findings

  • βEcd alone increased trabecular BV/TV by 41% at the 5th lumbar vertebral body and by 73% at the distal femur, with Tb.Th up 47% at the distal femur; it increased MS/BS by 43% and BFR/BS by 30% at the 4th lumbar vertebra, cortical Ct-BV by 4.5%, endocortical bone formation by 213%, and periosteal bone formation by 55%. Vertebral maximum load, ultimate stress, and toughness increased by 53%, 21%, and 16%; femoral values increased by 23%, 42%, and 9%, respectively.
  • GC reduced trabecular BV/TV by 32%, MS/BS by 46%, MAR by 23%, and BFR/BS by 60%; it decreased serum osteocalcin by 12% and increased serum CTX-1 by 125%. GC reduced vertebral maximum load, ultimate stress, and toughness by 24%, 19%, and 16%, reduced femoral cortical toughness by 24%, and reduced endocortical and periosteal bone formation by 99% and 120%, respectively.
  • GC + βEcd increased BV/TV by 43% versus GC, prevented the GC-induced inhibition of bone formation parameters, increased endocortical bone formation by 93% and periosteal bone formation by 59% versus GC, and increased vertebral maximum load, ultimate stress, and toughness by 44%, 31%, and 56% versus GC; femoral values increased by 16%, 18%, and 21%. βEcd did not prevent the loss in cortical bone volume.
  • In vitro, βEcd lowered pro-adipogenic gene expression (Cebp-α, Ppar-γ) by 5–20 fold and increased osteoblastic gene expression and CFU-Ob/CFU-F; dexamethasone decreased LC3+ primary osteoblast numbers by more than 50%, and βEcd prevented this. In vivo, GC reduced autophagy-associated genes including Atg7 and Beclin 1 by 1–5 fold, while βEcd co-treatment activated them by 5–10 fold; GC reduced Atg7 and Atg16L by more than 50%, and βEcd maintained these proteins and LC3II/I ratio near placebo levels.

Methods

  • Two-month-old male Swiss-Webster mice were randomized to four groups (abstract: n=8–10/group; methods: 8/group): placebo, placebo + βEcd (0.5 mg/kg, 5×/week), slow-release prednisolone (3.3 mg/kg/d), or prednisolone + βEcd for 21 days. Calcein was injected 7 and 2 days before euthanization.
  • Outcomes included serum hormones and bone turnover markers, microCT of the 5th lumbar vertebra and right femur, bone histomorphometry of lumbar vertebrae and femoral shafts, and biomechanical testing by vertebral compression and femoral three-point bending.
  • In vitro studies used bone marrow stromal cells for osteogenesis/adipogenesis assays and dsRed-LC3 reporter cells for autophagy; RNA from tibial cortical bone was analyzed by autophagy RT-PCR arrays, and Western blot measured Atg7, Atg16L, and LC3. Statistical analysis used group means/SDs and nonparametric Kruskal-Wallis tests.

Limitations

  • The paper states that only male mice were studied, at one time point (21 days), with a single βEcd dose regimen; female mice and dose-response studies were not performed. It also states the study only addressed prevention of GC-induced bone loss, not treatment of established chronic GC bone loss.
  • The paper states that young mice were used, long bone length was not measured, and growth effects of GC or βEcd could not be concluded; body mass changes may confound results, though the authors argue similar trends in weight-bearing and non-weight-bearing sites make this unlikely.
  • The paper notes that serum was collected only at one single time point and may not reflect dynamic osteoclast activity; βEcd had an inconsistent effect on resorption, inhibiting osteoclast maturation in vitro (data on file) while increasing serum CTX-1.

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

Integrity signals

Registry

Crossref · 10.1016/j.bone.2015.01.001

Cites

72 of 72 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 70 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    In vitro studies determined that beta-ecdysone prevented the glucocorticoid increase in autophagy of the bone marrow stromal cells.

    “In vitro studies determined that βEcd prevented the GC increase in autophagy of the bone marrow stromal cells”
    Quote in the paperSubstantial claim
  • Findingown data

    Body mass declined in the glucocorticoid and glucocorticoid plus beta-ecdysone groups by 20% during the three-week study period.

    “and declined in the GC and GC+ βEcd groups by 20% during this three-week study period (p < 0.05 vs. baseline).”
    Quote in the paperNot a substantial claim
  • Findingown data

    Concurrent treatment of beta-ecdysone and glucocorticoid completely prevented the glucocorticoid-induced reduction in trabecular bone volume.

    “Concurrent treatment of βEcd and GC completely prevented the GC-induced reduction in BFR, trabecular bone volume”
    Quote in the paperSubstantial claim
  • Findingown data

    Serum osteocalcin and CTX-1 levels increased significantly in the placebo plus beta-ecdysone group as compared to the placebo group.

    “Serum osteocalcin and CTX-1 levels increased significantly in PL + βEcd group as compared to PL group.”
    Quote in the paperNot a substantial claim
  • Findingown data

    Glucocorticoid treatment inhibited age-dependent trabecular gain and cortical bone expansion.

    “GC treatment inhibited age-dependent trabecular gain and cortical bone expansion”
    Quote in the paperSubstantial claim
  • Classified as probably not a substantial claim. 13
  • The quote only partly states this claim. 1
  • May combine two claims. 29

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

    Findingown data

    Body mass increased by approximately 25% in the placebo and placebo plus beta-ecdysone groups during the three-week study period.

    “Body mass increased by approximately 25% in the PL and PL + βEcd groups”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Findingown data

    In vitro studies determined that beta-ecdysone prevented the glucocorticoid increase in autophagy in whole bone.

    “as well as in whole bone.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Glucocorticoids non-significantly lowered trabecular bone volume/tissue volume and trabecular thickness in the distal femur.

    “GCs non-significantly lowered BV/TV and Tb.Th in the DFM”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Glucocorticoid plus beta-ecdysone had approximately 10% higher trabecular bone volume/tissue volume and trabecular number in the distal femur, but these changes were not significant compared to the glucocorticoid group.

    “GC+ βEcd had approximately 10% higher BV/TV and Th.N, but these changes were not significant compared to the GC group (Figure 1C).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Beta-ecdysone increased the differentiation of mesenchymal progenitor cells towards osteoblast in vitro, consistent with the observation of increasing the osteoblast surface following beta-ecdysone treatment in vivo.

    “Furthermore, our results demonstrated that βEcd increased the differentiation of mesenchymal progenitor cells towards osteoblast in vitro, consistent with the observation of increasing the osteoblast surface following βEcd treatment in vivo.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Glucocorticoid decreased LC3-II/I ratio by about 15%, suggesting LC3 conversion might be not significantly affected following glucocorticoid treatment in vivo.

    “On the other hand, GC decreased LC3-II/I ratio by about 15%, suggesting LC3 conversion might be not significantly affected following GC treatment in vivo.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Glucocorticoid reduced expression of Atg7 and Atg16L by more than 50%, indicating reduced formation of autophagosome.

    “GC reduced expression of Atg7 and Atg16L by more than 50%, indicating reduced formation of autophagosome.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Beta-ecdysone completely prevented losses in mechanical and material strengths in both vertebral trabecular bone and femoral cortical bone despite not completely preventing glucocorticoid-induced bone loss in these two bone sites.

    “On the other hand, βEcd completely prevented losses in mechanical (eg. Maximum load) and material strengths (stress and toughness) in both the vertebral trabecular bone and femoral cortical bone despite it did not completely prevent GC-induced bone loss in these two bone sites.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Beta-ecdysone might sustain the autophagic level in osteoblastic-like cells and in bone, whereas glucocorticoids suppressed the induction of autophagy.

    “Although far from conclusive, our exploratory experiments on autophagy in vitro and in vivo supported that βEcd might sustain the autophagic level in osteoblastic-like cells and in bone, whereas GC suppressed the induction of autophagy.”
    Quote in the paperSubstantial claim

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

    Findingnone given

    The mice used in the study gained about 20% body mass during the study, which might be similar to the skeleton of early adulthood finishing its maturation.

    “The mice we used in the study gained about 20% body mass during the study, which might be similar to the skeleton of early adulthood finishing its maturation.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Short-term administration of beta-ecdysone in growing male mice resulted in marked enhancements in both trabecular and cortical bone formation that were associated with significantly increased trabecular and cortical bone volume, both contributing to overall skeletal strength.

    “In summary, we have found that short-term administration of βEcd in growing male mice resulted in marked enhancements in both trabecular and cortical bone formation that were associated with significantly increased trabecular and cortical bone volume, both contributing to overall skeletal strength.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Glucocorticoid treatment reduced endosteal bone formation and periosteal bone expansion, while beta-ecdysone treatment prevented the detrimental effect of glucocorticoids on bone formation, especially on trabecular bone.

    “While GC treatment reduced endosteal bone formation and periosteal bone expansion, βEcd treatment prevented the detrimental effect of GC on bone formation, especially on trabecular bone.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Glucocorticoid use results in rapid trabecular bone loss and a high incident fracture risk.

    “However, GC use results in rapid trabecular bone loss and a high incident fracture risk [1, 2].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [1] Goemaere S Liberman UA Adachi JD Hawkins F Lane N Saag KG Schnitzer T Kaufman JM Malice MP Carofano W Daifotis A Incidence of nonvertebral fractures in relation to time on treatment and bone density in glucocorticoid-treated patients: a retrospective approach J Clin Rheumatol 2003 9 170 5 17041453 source

      Not checked in the free run.

    • cites [2] Silverman SL Lane NE Glucocorticoid-induced osteoporosis Curr Osteoporos Rep 2009 7 23 6 19239826 source

      Not checked in the free run.

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

    Premisecitation

    Beta-ecdysone increases muscle weight in rodents.

    “As it increases muscle weight in rodents [26-28], βEcd has been referred to as an “anabolic” naturally-derived supplement [29].”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [26] Cheng DM Kutzler LW Boler DD Drnevich J Killefer J Lila MA Continuous infusion of 20-hydroxyecdysone increased mass of triceps brachii in C57BL/6 mice Phytother Res 2013 27 107 11 22495969 PMC3410052 source

      Not checked in the free run.

    • cites [27] Esposito D Rathinasabapathy T Poulev A Komarnytsky S Raskin I Akt-dependent anabolic activity of natural and synthetic brassinosteroids in rat skeletal muscle cells J Med Chem 2011 54 4057 66 21491949 PMC3128125 source

      Not checked in the free run.

    • cites [28] Toth N Szabo A Kacsala P Heger J Zador E 20-Hydroxyecdysone increases fiber size in a muscle-specific fashion in rat Phytomedicine 2008 15 691 8 18585021 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    The use of bisphosphonates is not ideal in a growing skeleton because they inhibit bone remodeling and could increase the mineral in the bone matrix.

    “However, as bone is highly remodeled during childhood to maintain adequate mineralization of the rapidly growing skeleton, the use of BPs is not ideal as they inhibit bone remodeling and could increase the mineral in the bone matrix, which may not be ideal to use in a growing skeleton [16, 17].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [16] Roldan EJ Pasqualini T Plantalech L Bisphosphonates in children with osteogenesis imperfecta may improve bone mineralization but not bone strength. Report of two patients J Pediatr Endocrinol Metab 1999 12 555 9 10417973 source

      Not checked in the free run.

    • cites [17] Rudge S Hailwood S Horne A Lucas J Wu F Cundy T Effects of once-weekly oral alendronate on bone in children on glucocorticoid treatment Rheumatology (Oxford) 2005 44 813 8 15695300 source

      Not checked in the free run.

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

    Premisecitation

    Beta-ecdysone increases protein synthesis and reduces protein degradation in skeletal muscle cells.

    “It has been shown that βEcd increases protein synthesis and reduces protein degradation in the skeletal muscle cells [24, 25].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [24] Gorelick-Feldman J MacLean D Ilic N Poulev A Lila MA Cheng D Raskin I Phytoecdysteroids increase protein synthesis in skeletal muscle cells Journal of Agricultural and Food Chemistry 2008 56 3532 3537 18444661 source

      Not checked in the free run.

    • cites [25] Esposito D Komarnytsky S Shapses S Raskin I Anabolic effect of plant brassinosteroid FASEB J 2011 25 3708 19 21746867 PMC3177571 source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductcitation

    The beta-ecdysone dose was based on publications on myogenesis and the authors' in vitro experiments on osteogenesis and osteoclastogenesis using beta-ecdysone doses ranging from 10-3 to 10-9M.

    “The βEcd dose was based publications on myogenesis and our in vitro experiments on osteogenesis and osteoclastogenesis using βEcd doses ranging from 10-3 to 10-9M [25, 26].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [25] Esposito D Komarnytsky S Shapses S Raskin I Anabolic effect of plant brassinosteroid FASEB J 2011 25 3708 19 21746867 PMC3177571 source

      Not checked in the free run.

    • cites [26] Cheng DM Kutzler LW Boler DD Drnevich J Killefer J Lila MA Continuous infusion of 20-hydroxyecdysone increased mass of triceps brachii in C57BL/6 mice Phytother Res 2013 27 107 11 22495969 PMC3410052 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Beta-ecdysone stimulates mesenchymal stem cells' osteogenic differentiation and inhibits their adipogenic differentiation.

    “Additionally, βEcd is also found to stimulate mesenchymal stem cells' osteogenic differentiation but to inhibit their adipogenic differentiation [30].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [30] Gao L Cai G Shi X Beta-ecdysterone induces osteogenic differentiation in mouse mesenchymal stem cells and relieves osteoporosis Biol Pharm Bull 2008 31 2245 9 19043207 source

      Not checked in the free run.

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

    Study design and conductnone given

    Mice were randomized into 4 experimental groups of 8 animals each.

    “They were randomized into 4 experimental groups of 8 animals in each group.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Beta-ecdysone is reported to increase growth plate width in estrogen deficient rats and to have a marginal beneficial effect on trabecular bone and cartilage preserving following ovariectomy.

    “βEcd is reported to increase the growth plate width in estrogen deficient rats and to have a marginal beneficial effect on trabecular bone and cartilage preserving following ovariectomy (OVX) [31, 32].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [31] 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 17 350 5 20171072 source

      Not checked in the free run.

    • cites [32] Seidlova-Wuttke D Christel D Kapur P Nguyen BT Jarry H Wuttke W Beta-ecdysone has bone protective but no estrogenic effects in ovariectomized rats Phytomedicine 2010 17 884 9 20554186 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Beta-ecdysone prevented glucocorticoid-induced changes in bone formation, bone cell viability and bone mass.

    “In summary, βEcd prevented GC induced changes in bone formation, bone cell viability and bone mass.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Detrimental effects on both trabecular and cortical bone architecture and bone strength were apparent in mice receiving glucocorticoids, which included inhibitions in active bone formation at the trabecular bone and cortical bone expansion.

    “Detrimental effect on both trabecular and cortical bone architecture and bone strength were apparent in mice receiving GCs, which included inhibitions in active bone formation at the trabecular bone and cortical bone expansion.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Glucocorticoids potently suppress osteoblast activities and adversely affect bone mineralization.

    “As GCs potently suppress osteoblast activities and adversely affect bone mineralization [41, 46]”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [41] Yao W Cheng Z Pham A Busse C Zimmermann EA Ritchie RO Lane NE Glucocorticoid-induced bone loss in mice can be reversed by the actions of parathyroid hormone and risedronate on different pathways for bone formation and mineralization Arthritis Rheum 2008 58 3485 3497 18975341 PMC2597521 source

      Not checked in the free run.

    • cites [46] Balooch G Yao W Ager JW Balooch M Nalla RK Porter AE Ritchie RO Lane NE The aminobisphosphonate risedronate preserves localized mineral and material properties of bone in the presence of glucocorticoids Arthritis Rheum 2007 56 3726 3737 17968931 PMC6913894 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    The substantial improvements in bone formation in beta-ecdysone-treated mice translated into substantially higher bone mass with higher vertebral and cortical bone strength.

    “The substantial improvements in bone formation in βEcd-treated mice translated into substantially higher bone mass with higher vertebral and cortical bone strength.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    The anabolic agent hPTH (1-34) is superior to bisphosphonates in increasing spine and hip BMD and reduces the incidence of new vertebral fractures in glucocorticoid-induced osteoporosis populations for both men and women.

    “In support of this notion, it is reported that anabolic agent, hPTH (1-34), is superior to bisphosphonates in increasing spine and hip BMD and reduces the incidence of new vertebral fractures in glucocorticoid-induced osteoporosis (GIOP) populations for both men and women [11, 47-49].”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [11] Saag KG Shane E Boonen S Marin F Donley DW Taylor KA Dalsky GP Marcus R Teriparatide or alendronate in glucocorticoid-induced osteoporosis N Engl J Med 2007 357 2028 39 18003959 source

      Not checked in the free run.

    • cites [47] Devogelaer JP Sambrook P Reid DM Goemaere S Ish-Shalom S Collette J Su G Bucci-Rechtweg C Papanastasiou P Reginster JY Effect on bone turnover markers of once-yearly intravenous infusion of zoledronic acid versus daily oral risedronate in patients treated with glucocorticoids Rheumatology (Oxford) 2013 52 1058 69 23365149 source

      Not checked in the free run.

    • cites [48] Gluer CC Marin F Ringe JD Hawkins F Moricke R Papaioannu N Farahmand P Minisola S Martinez G Nolla JM Niedhart C Guanabens N Nuti R Martin-Mola E Thomasius F Kapetanos G Pena J Graeff C Petto H Sanz B Reisinger A Zysset PK Comparative effects of teriparatide and risedronate in glucocorticoid-induced osteoporosis in men: 18-month results of the EuroGIOPs trial J Bone Miner Res 2013 28 1355 68 23322362 PMC3708101 source

      Not checked in the free run.

    • cites [49] Saag KG Zanchetta JR Devogelaer JP Adler RA Eastell R See K Krege JH Krohn K Warner MR Effects of teriparatide versus alendronate for treating glucocorticoid-induced osteoporosis: thirty-six-month results of a randomized, double-blind, controlled trial Arthritis Rheum 2009 60 3346 55 19877063 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationcitation

    Beta-ecdysone treatment may activate bone turnover, increasing both bone resorption and bone formation, with the latter exceeding the former such that a net bone gain was observed.

    “Similar to PTH, βEcd treatment may activate bone turnover, increasing both bone resorption and bone formation, with the latter exceeding the former such that a net bone gain was observed [60].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [60] Hock JM Anabolic actions of PTH in the skeletons of animals J Musculoskelet Neuronal Interact 2001 2 33 47 15758475

      Not checked in the free run.

  • May combine two claims.

    Study design and conductown data

    Serum was collected only at one single time point (at the end of three months), so serum measurements may not reflect dynamic changes over the entire experiment on osteoclast activities.

    “However, serum was collected only at one single time point (eg, at the end of three months) and thus the serum measurements may not reflect the dynamic changes over the entire experiment on osteoclast activities.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Beta-ecdysone is reported to promote muscle cell growth via insulin and PI3K/Akt signaling.

    “Mechanistically, βEcd is reported to promote muscle cell growth via insulin and PI3K/Akt signaling and is reported to inhibit NF-kB activation in a cancer cell line [21, 25, 27, 61].”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [21] Badisco L Van Wielendaele P Vanden Broeck J Eat to reproduce: a key role for the insulin signaling pathway in adult insects Front Physiol 2013 4 202 23966944 PMC3735985 source

      Not checked in the free run.

    • cites [25] Esposito D Komarnytsky S Shapses S Raskin I Anabolic effect of plant brassinosteroid FASEB J 2011 25 3708 19 21746867 PMC3177571 source

      Not checked in the free run.

    • cites [27] Esposito D Rathinasabapathy T Poulev A Komarnytsky S Raskin I Akt-dependent anabolic activity of natural and synthetic brassinosteroids in rat skeletal muscle cells J Med Chem 2011 54 4057 66 21491949 PMC3128125 source

      Not checked in the free run.

    • cites [61] Peschel W Kump A Prieto JM Effects of 20-hydroxyecdysone, Leuzea carthamoides extracts, dexamethasone and their combinations on the NF-kappaB activation in HeLa cells J Pharm Pharmacol 2011 63 1483 95 21988429 source

      Not checked in the free run.

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

    Premisecitation

    Beta-ecdysone is reported to inhibit NF-kB activation in a cancer cell line.

    “Mechanistically, βEcd is reported to promote muscle cell growth via insulin and PI3K/Akt signaling and is reported to inhibit NF-kB activation in a cancer cell line [21, 25, 27, 61].”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [21] Badisco L Van Wielendaele P Vanden Broeck J Eat to reproduce: a key role for the insulin signaling pathway in adult insects Front Physiol 2013 4 202 23966944 PMC3735985 source

      Not checked in the free run.

    • cites [25] Esposito D Komarnytsky S Shapses S Raskin I Anabolic effect of plant brassinosteroid FASEB J 2011 25 3708 19 21746867 PMC3177571 source

      Not checked in the free run.

    • cites [27] Esposito D Rathinasabapathy T Poulev A Komarnytsky S Raskin I Akt-dependent anabolic activity of natural and synthetic brassinosteroids in rat skeletal muscle cells J Med Chem 2011 54 4057 66 21491949 PMC3128125 source

      Not checked in the free run.

    • cites [61] Peschel W Kump A Prieto JM Effects of 20-hydroxyecdysone, Leuzea carthamoides extracts, dexamethasone and their combinations on the NF-kappaB activation in HeLa cells J Pharm Pharmacol 2011 63 1483 95 21988429 source

      Not checked in the free run.

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

    Premisecitation

    Beta-ecdysone binds to ecdysone receptor with EC50 of 0.3 nM, a ligand-activated transcriptional factor found in arthropods.

    “βEcd binds to ecdysone receptor with EC50 of 0.3nm, a ligand-activated transcriptional factor found in arthropods [62, 63].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [62] Riddiford LM Cherbas P Truman JW Ecdysone receptors and their biological actions Vitam Horm 2000 60 1 73 11037621 source

      Not checked in the free run.

    • cites [63] Lapenna S Friz J Barlow A Palli SR Dinan L Hormann RE Ecdysteroid ligand-receptor selectivity--exploring trends to design orthogonal gene switches FEBS J 2008 275 5785 809 19021756 source

      Not checked in the free run.

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

    Premisecitation

    Endogenous glucocorticoids are the main hormones released in response to stress and glucocorticoid excess has been associated with nutrition deprivation and accelerating aging process.

    “Endogenous GCs are the main hormones released in response to stress and GC excess has been associated with nutrition deprived and accelerating aging process [66, 67].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [66] Zheng B Ohkawa S Li H Roberts-Wilson TK Price SR FOXO3a mediates signaling crosstalk that coordinates ubiquitin and atrogin-1/MAFbx expression during glucocorticoid-induced skeletal muscle atrophy FASEB J 2010 24 2660 9 20371624 PMC2909288 source

      Not checked in the free run.

    • cites [67] Sapolsky RM Krey LC McEwen BS Prolonged glucocorticoid exposure reduces hippocampal neuron number: implications for aging J Neurosci 1985 5 1222 7 3998818 PMC6565052 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    The stress from low dose glucocorticoids is nontoxic, allowing the cells to respond to the initial cellular or nutritional insults, which become no longer effective when the glucocorticoid dose is high or with prolonged treatment.

    “The stress from low dose GC is nontoxic, allowing the cells to respond to the initial cellular or nutritional insults, which become no longer effective when the GC dose is high or with prolonged treatment.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Autophagy is not a simple on-off phenomenon; the level of autophagy is maintained at a fine equilibrium.

  • Classified as probably not a substantial claim. May combine two claims.

    Study design and conductown data

    The study used relatively young mice, and the effect of glucocorticoids and beta-ecdysone on bone growth inhibition versus a true decrease in bone volume would need careful dissection.

    “We used relatedly young mice that we would have to carefully dissect out the effect of GC and βEcd on bone growth inhibition verse a true decrease in bone volume.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    Only male mice were studied at one time point (21 days), with a single dose regimen of beta-ecdysone.

    “First, only male mice were studied one time point (21 days), with a single dose regimen of βEcd.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Study design and conductown data

    The study did not measure the length of long bones and could not conclude if glucocorticoids or beta-ecdysone affect growth.

    “We did not measure the length of long bones and could not conclude if GC or βEcd affect growth.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Osteocyte autophagy is crucial to the regulation of bone structure and plays a role in glucocorticoid-induced bone fragility.

    “We have reported that osteocyte autophagy is crucial to the regulation of bone structure and plays a role in GC induced bone fragility [44, 68].”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [44] Jia J Yao W Guan M Dai W Shahnazari M Kar R Bonewald L Jiang JX Lane NE Glucocorticoid dose determines osteocyte cell fate FASEB J 2011 25 3366 76 21705669 PMC3177583 source

      Not checked in the free run.

    • cites [68] Xia X Kar R Gluhak-Heinrich J Yao W Lane NE Bonewald LF Biswas SK Lo WK Jiang JX Glucocorticoid induced autophagy in osteocytes J Bone Miner Res 2010 PMC3179284 20564240 source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    This study only addressed the effect of beta-ecdysone for the prevention of glucocorticoid-induced bone loss.

    “Second, this study only addressed the effect of βEcd for the prevention of GC induced bone loss.”
    Quote in the paperSubstantial claim
37 of 37

  • The paper relies on this without support.

    The mouse model of prednisolone-induced bone loss is representative of glucocorticoid-induced bone loss in humans.

    “Since GC use in children often results in growth retardation, [6, 7, 33] through GC induced inhibition of osteoblasts through multiple mechanisms [34], we seek to determine if βEcd can rescue the GC-suppression on bone formation.”

  • The paper relies on this without support.

    Autophagy is not a simple on-off phenomenon; the level of autophagy is maintained at a fine equilibrium.

    “Autophagy is not a simple on – off phenomenon allowing a quick judgment of good/protective or bad/detrimental effect on cell metabolism. Rather, the level of autophagy has been maintained at a fine equilibrium so if or when the stress-inducing agent is removed, the cells might go on to survive.”

  • The paper relies on this without support.

    Beta-ecdysone is structurally similar to testosterone and is considered to have a steroid hormone-like effect.

    “Since βEcd is structurally similar to testosterone, it is considered to have a steroid hormone-like effect.”
3 of 3

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 alone increased bone formation, and when added concurrently with glucocorticoid, it compensated for the detrimental effect and prevented GC-induced autophagy in bone marrow stromal cells.

    “Moreover, 20E alone increased bone formation, and when added concurrently with GC, it compensated for the detrimental effect and prevented the GC-induced autophagy in bone marrow stromal cells, making it a promising neutralizer of GC side effects.”

    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 [32]

    The Results (bone volume and bone turnover changes; page unavailable) report that β-ecdysone alone increased bone formation and concurrent treatment prevented GC-induced reductions. But the Results (effect on osteogenesis and autophagy; page unavailable) report the opposite autophagy direction: dexamethasone decreased LC3-positive osteoblasts derived from bone marrow stromal cells, and β-ecdysone prevented that decrease—not GC-induced autophagy. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • GC effect on cellular autophagy reverses between abstract and results

    Major

    The abstract describes a GC-induced increase in autophagy that βEcd prevented. The results instead report that Dex decreased the proportion of LC3-positive osteoblasts and βEcd prevented that decrease.

  • Abstract understates the reported cortical bone-formation losses

    Major

    The abstract says glucocorticoids lowered bone formation rate by 30-50% at both cortical surfaces, whereas the results report reductions of 99% at the endocortical surface and 120% at the periosteal surface.

  • Cortical bone-loss prevention is described oppositely

    Major

    The abstract says βEcd partially prevented cortical bone loss, but the cortical-bone results say it did not prevent the loss in cortical bone volume.

  • Serum collection timing conflicts with study duration

    Major

    The discussion places the single serum collection at the end of three months, although the methods say the mice were sacrificed after three weeks of treatment.

  • βEcd dosing frequency differs between abstract and methods

    Minor

    The abstract specifies βEcd at 0.5 mg/kg per day, while the methods specify 0.5 mg/kg five times per week rather than daily.

  • Reported 120% reduction implies a negative bone-formation rate

    Minor

    A 120% reduction from the placebo bone-formation rate would make the GC group's rate negative. That conflicts with the paper's definition of the rate as a product of mineralizing surface and mineral apposition rate, quantities that cannot be negative.

6 of 6

Cites

Flagged first, then by citations

Research works printed but not linked: 1

  1. Yao, 2010, Overexpression of secreted frizzled-related protein 1 inhibits bone formation and attenuates parathyroid hormone bone anabolic effects, J Bone Miner Res, 25, 190, 10.1359/jbmr.090719 · DOI

Cited by

Sorted by citations

Related

Authors
Dai, W., Jiang, L., Lay, Y.-A. E., Chen, H., Jin, G., Zhang, H., Kot, A., Ritchie, R. O., Lane, N. E., & Yao, W.
Affiliations
  • Longhua Hospital Shanghai University of Traditional Chinese Medicine (CN)
  • University of California Davis Medical Center (US)
  • Shanghai University of Traditional Chinese Medicine (CN)
  • University of California, Berkeley (US)
Corresponding author
Yao, Wei
ORCID
Venue
Bone
Publisher
Elsevier BV
Published
2015-05
Volume
74
Pages
48-57
ISSN
  • 1873-2763 (ISSN-L)
  • 8756-3282
DOI
10.1016/j.bone.2015.01.001
PMC
PMC4355031
OpenAlex
W2125913965
Cited by
30
References
72
Citations by year
2025: 2 · 2024: 1 · 2023: 1 · 2022: 4 · 2021: 6 · 2020: 2 · 2019: 2 · 2018: 3 · 2017: 5 · 2016: 1 · 2015: 3
FWCI
4.12
Type
journal article
Language
en
Open access
Licence
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations
  • Bone (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
National Institute of Arthritis and Musculoskeletal and Skin Diseases (R01-AR 43052, R01AR043052, P50AR063043, K24AR048841, R01AR061366)
Topics
  • Health Sciences › Medicine › Orthopedics and Sports Medicine › Bone health and osteoporosis research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bone Metabolism and Diseases
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Medicinal Plant Pharmacodynamics Research
Keywords
20-hydroxyecdysone, phytoecdysteroids, glucocorticoid-induced osteoporosis, prednisolone, bone formation rate, trabecular bone volume, cortical bone loss, mesenchymal stem cells, autophagy, bone formation, bone mass
MeSH terms
Animals, Autophagy, Biomechanical Phenomena, Body Weight, Bone and Bones, Cell Differentiation, Ecdysterone, Glucocorticoids, Hormones, Male, Organ Size, Osteoblasts, Osteogenesis, Bone Remodeling, Mice, X-Ray Microtomography, Mesenchymal Stem Cells

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-10.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-10.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

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