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Cited by 13Cholesterol and Lipid MetabolismLiver Disease Diagnosis and TreatmentLipid metabolism and disorders

Effect of Ecdysterone on the Hepatic Transcriptome and Lipid Metabolism in Lean and Obese Zucker Rats

Open access
Marschall, Magdalena J. M., Ringseis, Robert, Gessner, Denise K., Grundmann, Sarah M., Most, Erika, Wen, Gaiping, Maheshwari, Garima, Zorn, Holger +1 more·2021·International Journal of Molecular Sciences
DOI 10.3390/ijms22105241PMC PMC8156757
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Abstract

Conflicting reports exist with regard to the effect of ecdysterone, the predominating representative of steroid hormones in insects and plants, on hepatic and plasma lipid concentrations in different rodent models of obesity, fatty liver, and diabetes, indicating that the effect is dependent on the rodent model used. Here, the hypothesis was tested for the first time that ecdysterone causes lipid-lowering effects in genetically obese Zucker rats. To test this hypothesis, two groups of male obese Zucker rats (n = 8) were fed a nutrient-adequate diet supplemented without or with 0.5 g ecdysterone per kg diet. To study further if ecdysterone is capable of alleviating the strong lipid-synthetic activity in the liver of obese Zucker rats, the study included also two groups of male lean Zucker rats (n = 8) which also received either the ecdysterone-supplemented or the non-supplemented diet. While hepatic and plasma concentrations of triglycerides and cholesterol were markedly higher in the obese compared to the lean rats (p < 0.05), hepatic and plasma triglyceride and cholesterol concentrations did not differ between rats of the same genotype fed the diets without or with ecdysterone. In conclusion, the present study clearly shows that ecdysterone supplementation does not exhibit lipid-lowering actions in the liver and plasma of lean and obese Zucker rats.

AI snapshot

Written from the paper’s full text.

TL;DR

Ecdysterone supplementation did not lower hepatic or plasma lipids in lean or obese Zucker rats. The study rejects the hypothesis that ecdysterone causes lipid-lowering effects in this genetic obesity model.

Key findings

  • Hepatic and plasma triglyceride and cholesterol concentrations were higher in obese than lean rats (p < 0.05), but did not differ between rats of the same genotype fed diets without or with 0.5 g ecdysterone/kg diet.
  • Ecdysterone had only weak effects on the hepatic transcriptome: in lean rats, 414 transcripts were differentially expressed (208 upregulated, 206 downregulated), but only Inmt was upregulated >2.0-fold and only Tsku was downregulated <−2.0-fold; in obese rats, 684 transcripts were differentially expressed (369 upregulated, 315 downregulated), with only six upregulated >2.0-fold and one downregulated <−2.0-fold.
  • Among 32 lipid-synthetic genes upregulated 1.56- to 44.5-fold in obese vs. lean rats, only Fasn was slightly downregulated (−1.39-fold) by ecdysterone in obese rats.
  • Plasma fructosamine was higher in obese than lean rats (p < 0.05) but was not affected by ecdysterone; muscle weights and M. rectus femoris morphology were also not affected by ecdysterone.

Methods

  • Male 25-week-old obese (fa/fa) and lean (fa/+) Zucker rats were assigned to four groups of n = 8: lean control, lean ecdysterone, obese control, obese ecdysterone.
  • Rats received a nutrient-adequate basal diet without or with 0.5 g ecdysterone/kg diet for 4 weeks, providing approximately 20 mg ecdysterone/kg body weight/day.
  • Outcomes included hepatic and plasma triglyceride and cholesterol, hepatic fatty acid concentrations, hepatic transcriptome by microarray with qPCR validation, plasma fructosamine, and histological analysis of liver and M. rectus femoris; statistical analysis used two-way ANOVA with Tukey test where applicable.

Limitations

  • The paper states that adipose tissue depots were not excised, so future studies should evaluate whether ecdysterone affects adipose tissue metabolism in Zucker rats.
  • The paper notes that the ecdysterone effect on hepatic transcriptome was weak and that false discovery rate adjustment failed because adjusted p-values for all transcripts were >0.05; differential expression therefore used FC > 1.3 or < −1.3 with p < 0.05.
  • Evident from the text: the study used a single ecdysterone dose, a 4-week treatment period, and 25-week-old rats with already-developed steatosis and hyperlipidaemia, so dose-, duration-, and age-dependent effects cannot be excluded.

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

Integrity signals

Registry

Crossref · 10.3390/ijms22105241

Cites

40 of 40 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 22 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Hepatic and plasma concentrations of triglycerides and cholesterol were higher in obese Zucker rats than in lean Zucker rats (p < 0.05).

    “While hepatic and plasma concentrations of triglycerides and cholesterol were markedly higher in the obese compared to the lean rats (p < 0.05)”
    • Also stated in: § 2.2. Hepatic and Plasma Lipid Concentrations · plain
      “the obese rats had higher concentrations of triglycerides and cholesterol in liver and plasma than the lean rats (p < 0.05).”
    Quote in the paperSubstantial claim
  • Findingown data

    The most enriched KEGG pathways assigned to transcripts upregulated in OC vs. LC predominantly comprised lipid metabolic pathways, including fatty acid metabolism, biosynthesis of unsaturated fatty acids, fatty acid elongation, fatty acid degradation, and PPAR signalling pathway.

    “The most enriched KEGG pathways assigned to the transcripts upregulated in group OC vs. LC predominantly comprised lipid metabolic pathways, such as fatty acid metabolism, biosynthesis of unsaturated fatty acids, fatty acid elongation, fatty acid degradation, and PPAR signalling pathway (Figure 3b).”
    Quote in the paperNot a substantial claim
  • Findingown data

    Hepatic concentrations of 20:4 n-6 and 22:6 n-3 were lower in obese than lean Zucker rats.

    “those of 20:4 n-6 and 22:6 n-3 were lower in obese rats than in lean rats.”
    Quote in the paperNot a substantial claim
  • Findingown data

    Hepatic concentrations of most individual fatty acids (14:0, 14:1 n-5, 16:0, 16:1 n-7, 18:0, 18:1 n-9, 18:3 n-3, 18:3 n-6, 20:3 n-6) and the sum of all individual fatty acids were higher in obese than lean Zucker rats.

    “Hepatic concentrations of most individual fatty acids (14:0, 14:1 n-5, 16:0, 16:1 n-7, 18:0, 18:1 n-9, 18:3 n-3, 18:3 n-6, 20:3 n-6) and the sum of all individual fatty acids were higher and those of 20:4 n-6 and 22:6 n-3 were lower in obese rats than in lean rats.”
    Quote in the paperNot a substantial claim
  • Findingown data

    Hepatic and plasma triglyceride and cholesterol concentrations did not differ between Zucker rats of the same genotype fed diets without or with ecdysterone.

    “hepatic and plasma triglyceride and cholesterol concentrations did not differ between rats of the same genotype fed the diets without or with ecdysterone.”
    • Also stated in: § 2.2. Hepatic and Plasma Lipid Concentrations · plain
      “Liver and plasma triglyceride and cholesterol concentrations of the rats were influenced by the genotype but not by ecdysterone (Figure 1a)”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The study had sufficient statistical power to detect a lipid-lowering effect of ecdysterone in Zucker rats.
  • May combine two claims. 13
  • Classified as probably not a substantial claim. 23

  • May combine two claims.

    Findingown data

    For all validated transcripts, the direction of effect (positive or negative FC) was the same between microarray and qPCR, whereas effect size differed to some extent.

    “the effect direction (positive or negative FC) was the same between microarray and qPCR for all validated transcripts, whereas the effect size (value of FC) differed to some extent for the validated transcripts between microarray and qPCR.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Comparing obese and lean Zucker rats fed basal diet without ecdysterone (OC vs. LC), 1981 hepatic transcripts were differentially regulated (FC > 1.3 or < -1.3, p < 0.05), with 1092 upregulated and 889 downregulated in OC vs. LC.

    “When comparing the obese and the lean groups of rats fed the basal diet without ecdysterone (OC vs. LC), a total of 1981 hepatic transcripts were differentially regulated according to the two-filter criteria (FC > 1.3 or < −1.3, p < 0.05); of those, 1092 transcripts were upregulated and 889 transcripts were downregulated in the OC group, compared to the LC group (Figure 2a).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    qPCR analysis revealed 17 of the validated transcripts were regulated significantly (p < 0.05), whereas two (Srebf1, Ldlr) were not regulated (p > 0.05).

    “Statistical analysis of qPCR data revealed that 17 (G6pd, Scd2, Elovl6, Gpam, Cd36, Me1, Fasn, Fads2, Elovl5, Car3, Dhrs7, Sult1c3, Nrep, Cyp3a18, Sds, Cdh17, Acnat2) of the validated transcripts were regulated significantly (p < 0.05), whereas two (Srebf1, Ldlr) of the transcripts were not regulated (p > 0.05).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Among upregulated transcripts in OE vs. OC, only six genes were regulated > 2.0-fold and the strongest ecdysterone-induced gene was Gadd45a (2.57).

    “Amongst the upregulated genes, only six genes were regulated >2.0-fold and the strongest ecdysterone-induced gene was Gadd45a (2.57).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    No difference in liver lipid accumulation or morphology was observed between lean rats fed with or without ecdysterone and between obese rats fed with or without ecdysterone.

    “No difference was observed between lean rats fed with (LE) or without ecdysterone (LC) and between obese rats fed with (OE) or without ecdysterone (OC).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Initial and final body weights, body weight gain, feed intake, feed:gain ratio, and organ weights of the rats were influenced by genotype but not by ecdysterone.

    “Initial and final body weights, body weight gain, feed intake, feed:gain ratio, and organ weights of the rats were only influenced by the genotype but not by ecdysterone (Table 1).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Oil Red O-stained liver sections of lean Zucker rats showed normal parenchyma structure and liver cell morphology with no abnormalities.

    “the Oil Red O-stained liver sections of the two lean groups (LC, LE) showed a normal appearance of the parenchyma structure with normal liver cell morphology, clear edges, clearly visible haematoxylin-stained nuclei, and no abnormalities (Figure 1b).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The top 10 downregulated transcripts in OE vs. OC were Angptl8 (-2.19), Trim24 (-1.98), Vom2r75 (-1.86), Ces4a (-1.78), Aqp11 (-1.76), Gstt3 (-1.74), Map3k15 (-1.73), Enpp5 (-1.72), Hebp2 (-1.71), and RGD1559459 (-1.69).

    “The top 10 downregulated transcripts were in increasing order of their FCs (in brackets) as follows: Angptl8 (−2.19), Trim24 (−1.98), Vom2r75 (−1.86), Ces4a (−1.78), Aqp11 (−1.76), Gstt3 (−1.74), Map3k15 (−1.73), Enpp5 (−1.72), Hebp2 (−1.71), and RGD1559459 (−1.69).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Obese Zucker rats had higher initial and final body weights, body weight gain, feed intake, weights of liver, heart and kidneys, and lower weights of selected muscles than lean Zucker rats (p < 0.05).

    “Obese rats had higher initial and final body weights, body weight gain, feed intake, weights of liver, heart and kidneys, and lower weights of selected muscles than lean rats (p < 0.05).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Ecdysterone supplementation did not affect the weights of different muscles excised (M. rectus femoris, M. gastrocnemius, M. soleus, M. vastus intermedius, M. vastus medialis) or muscle morphology in Zucker rats.

    “the weights of different muscles excised, such as M. rectus femoris, M. gastrocnemius, M. soleus, M. vastus intermedius and M. vastus medialis, and muscle morphology were not affected by ecdysterone supplementation.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Ecdysterone supplemented in the diet at 0.5 g/kg is absorbed and reaches the liver and skeletal muscle at bioactive concentrations.
    • Rests on an assumption: The 4-week supplementation period is sufficient to detect effects on hepatic transcriptome and lipid metabolism if they exist.
    • Rests on an assumption: The Zucker rat model is a valid model for human obesity and metabolic syndrome.

  • May combine two claims.

    Findingown data

    GSEA of transcripts upregulated in LE vs. LC revealed only two enriched (p < 0.05) biological process terms, membrane depolarisation during action potential and sodium ion transport, and only one enriched KEGG pathway, carbohydrate digestion and absorption.

    “GSEA of the transcripts upregulated in group LE vs. LC revealed only two enriched (p < 0.05) biological process terms, namely, membrane depolarisation during action potential and sodium ion transport (Figure 5a), and only one enriched (p < 0.05) KEGG pathway, i.e., carbohydrate digestion and absorption.”
    Quote in the paperNot a substantial claim

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

    Interpretationcitation

    Ecdysteroids act on the PI3K pathway.

    “The observations from C2C12 cell incubations that ecdysterone does not bind to the androgen receptor, but the protein anabolic effects of ecdysterone are completely abolished by a PI3K inhibitor suggest that ecdysteroids act on the PI3K pathway which is known to promote skeletal muscle growth [25].”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    The PI3K pathway is known to promote skeletal muscle growth.

    “which is known to promote skeletal muscle growth [25].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [25] Guttridge, 2004, Signaling pathways weigh in on decisions to make or break skeletal muscle, Curr. Opin. Clin. Nutr. Metab. Care source

      Not checked in the free run.

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

    Premisecitation

    Ecdysteroids exert anabolic effects in a wide variety of vertebrates, such as mice, rats, pigs, and Japanese quails.

    “Evidence from several earlier studies exists that ecdysteroids exert anabolic effects in a wide variety of vertebrates, such as mice [19,20], rats [21], pigs [22], and Japanese quails [23].”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [19] Stopka, 1999, Effect of insect hormone, 20-hydroxyecdysone on growth and reproduction in mice, Acta Soc. Zoo. Bohem.

      Not checked in the free run.

    • cites [20] Todorov, 2000, The effect of ecdysterone on the biosynthesis of proteins and nucleic acids in mice, Pharm. Chem. J. source

      Not checked in the free run.

    • cites [21] Syrov, 1976, On the anabolic activity of the phytoecdysone-ecdysterone isolated from Rhaponticum carthamoides (Willd.) Iljin, Farmakol. Toksikol.

      Not checked in the free run.

    • cites [22] Kratky, 1997, Effect of 20-hydroxyecdysone on the protein synthesis of pigs, Zivocisna Vyrob.

      Not checked in the free run.

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

    Premisecitation

    Ecdysteroids, including ecdysterone, increase protein synthesis in C2C12 myotubes.

    “it has been demonstrated that ecdysteroids, including ecdysterone, increase protein synthesis in C2C12 myotubes [24].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [24] Maclean, 2008, Phytoecdysteroids increase protein synthesis in skeletal muscle cells, J. Agric. Food Chem. source

      Not checked in the free run.

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

    Premisecitation

    Ecdysterone does not bind to the androgen receptor.

    “The observations from C2C12 cell incubations that ecdysterone does not bind to the androgen receptor, but the protein anabolic effects of ecdysterone are completely abolished by a PI3K inhibitor suggest that ecdysteroids act on the PI3K pathway which is known to promote skeletal muscle growth [25].”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Daily administration of ecdysterone (50 mg/kg body weight) via gavage for 4 weeks increases front limb grip strength of rats.

    “Gorelick-Feldman et al. [24] showed that daily administration of ecdysterone (50 mg/kg body weight) via gavage for 4 weeks increases front limb grip strength of rats indicating that the protein anabolic effect of ecdysterone translates into improved physical performance.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [24] Maclean, 2008, Phytoecdysteroids increase protein synthesis in skeletal muscle cells, J. Agric. Food Chem. source

      Not checked in the free run.

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

    Premisecitation

    The protein anabolic effects of ecdysterone are completely abolished by a PI3K inhibitor.

    “The observations from C2C12 cell incubations that ecdysterone does not bind to the androgen receptor, but the protein anabolic effects of ecdysterone are completely abolished by a PI3K inhibitor suggest that ecdysteroids act on the PI3K pathway which is known to promote skeletal muscle growth [25].”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Anti-inflammatory interventions are capable of attenuating deleterious skeletal muscle changes in obese Zucker rats by inhibiting inflammatory signalling pathways in skeletal muscle.

    “The observation that anti-inflammatory interventions are capable of attenuating these deleterious skeletal muscle changes in obese Zucker rats by inhibiting inflammatory signalling pathways in skeletal muscle [29]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [29] Acevedo, L.M., Raya, A.I., Martínez-Moreno, J.M., Aguilera-Tejero, E., and Rivero, J.L. (2017). Mangiferin protects against adverse skeletal muscle changes and enhances muscle oxidative capacity in obese rats. PLoS ONE, 12. source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Lean rats were randomly assigned to two groups (lean control and lean ecdysterone) of eight rats each.

    “The lean rats were randomly assigned to two groups [lean control (LC), lean ecdysterone (LE)] of eight rats each.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductown data

    RNA quality was checked (A260:A280 ratios and RNA integrity number values).

    “After checking RNA quality [A260:A280 ratios and RNA integrity number values were 1.84 ± 0.05 (mean ± SD) and 7.4 ± 0.6, respectively], as described in [37]”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [37] Gessner, D.K., Winkler, A., Koch, C., Dusel, G., Liebisch, G., Ringseis, R., and Eder, K. (2017). Analysis of hepatic transcript profile and plasma lipid profile in early lactating dairy cows fed grape seed and grape marc meal extract. BMC Genom., 18. source

      Not checked in the free run.

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

    Study design and conductown data

    The animal experiment was approved by the local Animal Care and Use Committee.

    “The animal experiment was approved by the local Animal Care and Use Committee (Permission no. and date: JLU 725_M, 16 September 2019).”
    Quote in the paperNot a substantial claim

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

    Study design and conductcitation

    Normalisation of qPCR data was carried out according to Vandesompele et al. using the three most stable reference genes (Actb, Atp5b, Canx) out of seven potential reference genes tested.

    “Normalisation of qPCR data was carried out according to Vandesompele et al. [43] using the three most stable (Actb, Atp5b, Canx) out of seven potential reference genes tested.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [43] Vandesompele, 2002, Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes, Genome Biol. source

      Not checked in the free run.

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

    Study design and conductcitation

    Microarray data have been deposited in MIAME compliant format in NCBI's Gene Expression Omnibus public repository (GEO accession no. GSE168390).

    “The microarray data have been deposited in MIAME compliant format in the NCBI’s Gene Expression Omnibus public repository ([38]; GEO accession no. GSE168390).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [38] Edgar, 2002, Gene Expression Omnibus: NCBI gene expression and hybridization array data repository, Nucleic Acids Res. source

      Not checked in the free run.

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

    Premisecitation

    A marked inhibition of lipid synthetic pathways in the liver is an important mechanism underlying the lipid-lowering action of Tenebrio molitor meal.

    “As an important mechanism underlying this lipid-lowering action of Tenebrio molitor meal, a marked inhibition of lipid synthetic pathways in the liver has been identified [17,18].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [17] Gessner, 2019, Insect Meal as Alternative Protein Source Exerts Pronounced Lipid-Lowering Effects in Hyperlipidemic Obese Zucker Rats, J. Nutr. source

      Not checked in the free run.

    • cites [18] Meyer, 2019, The Antisteatotic and Hypolipidemic Effect of Insect Meal in Obese Zucker Rats is Accompanied by Profound Changes in Hepatic Phospholipid and 1-Carbon Metabolism, Mol. Nutr. Food Res. source

      Not checked in the free run.

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

    Study design and conductown data

    Microarray data of selected differentially expressed transcripts between groups OC and LC were validated by qPCR.

    “Microarray data of selected differentially expressed transcripts between groups OC and LC were validated by qPCR.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    The ecdysterone used was chemically characterised by HPLC and mass spectrometry.

    “The chemical characterisation of the aforementioned ecdysterone was carried out by subjecting it to high-performance liquid chromatography (HPLC) according to the method described by Sreejit et al. [32], with slight modifications, and mass spectrometry (MS).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [32] Sreejit, 2019, Isolation, quantification and chemical characterization of ecdysterone from medicinal plants of Kerala, Western Ghats, J. Pharmacogn. Phytochem.

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Ecdysterone has neither anabolic nor anti-catabolic effects on skeletal muscle in Zucker rats.

    “Our findings, therefore, suggest that ecdysterone has neither anabolic nor anti-catabolic effects on skeletal muscle in Zucker rats.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Ecdysterone supplemented in the diet at 0.5 g/kg is absorbed and reaches the liver and skeletal muscle at bioactive concentrations.
    • Rests on an assumption: The 4-week supplementation period is sufficient to detect effects on hepatic transcriptome and lipid metabolism if they exist.
    • Rests on an assumption: The Zucker rat model is a valid model for human obesity and metabolic syndrome.

  • Classified as probably not a substantial claim.

    Study design and conductown data

    Obese rats were randomly assigned to two groups (obese control and obese ecdysterone) of eight rats each.

    “Additionally, the obese rats were randomly assigned to two groups [obese control (OC), obese ecdysterone (OE)] of eight rats each.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Feeding of protein-rich insect meal produced from Tenebrio molitor markedly decreases liver and/or plasma lipids in obese Zucker rats.

    “recent reports showed that feeding of protein-rich insect meal produced from industrialised mass-rearing of the edible species Tenebrio molitor markedly decreases liver and/or plasma lipids in obese Zucker rats [17,18,30].”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [17] Gessner, 2019, Insect Meal as Alternative Protein Source Exerts Pronounced Lipid-Lowering Effects in Hyperlipidemic Obese Zucker Rats, J. Nutr. source

      Not checked in the free run.

    • cites [18] Meyer, 2019, The Antisteatotic and Hypolipidemic Effect of Insect Meal in Obese Zucker Rats is Accompanied by Profound Changes in Hepatic Phospholipid and 1-Carbon Metabolism, Mol. Nutr. Food Res. source

      Not checked in the free run.

    • cites [30] Meyer, S., Schäfer, L., Röhrig, J., Maheshwari, G., Most, E., Zorn, H., Ringseis, R., Eder, K., and Gessner, D.K. (2020). Supplementation of Sulfur-Containing Amino Acids or Essential Amino Acids Does Not Reverse the Hepatic Lipid-Lowering Effect of a Protein-Rich Insect Meal in Obese Zucker Rats. Nutrients, 12. source

      Not checked in the free run.

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

    Premisecitation

    The characteristically low methionine concentration of insect meal or a decreased cysteine synthesis secondary to a reduced methionine availability are not causative for the lipid-lowering effects of insect meal in obese Zucker rats.

    “we have shown recently that the characteristically low methionine concentration of insect meal or a decreased cysteine synthesis secondary to a reduced methionine availability are not causative [30]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [30] Meyer, S., Schäfer, L., Röhrig, J., Maheshwari, G., Most, E., Zorn, H., Ringseis, R., Eder, K., and Gessner, D.K. (2020). Supplementation of Sulfur-Containing Amino Acids or Essential Amino Acids Does Not Reverse the Hepatic Lipid-Lowering Effect of a Protein-Rich Insect Meal in Obese Zucker Rats. Nutrients, 12. source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    All data were checked for normality distribution by an Anderson-Darling test.

    “All data were checked for normality distribution by an Anderson–Darling test.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductreasoning

    The filter criteria FC > 1.3 and p < 0.05 were used for defining differentially expressed transcripts.

    “transcripts with an FC > 1.3 and a Student’s t-test p-value < 0.05 were defined as upregulated, and those with aN FC < −1.3 and a Student’s t-test p-value < 0.05 as downregulated for the comparison OE vs. OC and LE vs. LC.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [17] Gessner, 2019, Insect Meal as Alternative Protein Source Exerts Pronounced Lipid-Lowering Effects in Hyperlipidemic Obese Zucker Rats, J. Nutr. source

      Not checked in the free run.

    • cites [37] Gessner, D.K., Winkler, A., Koch, C., Dusel, G., Liebisch, G., Ringseis, R., and Eder, K. (2017). Analysis of hepatic transcript profile and plasma lipid profile in early lactating dairy cows fed grape seed and grape marc meal extract. BMC Genom., 18. source

      Not checked in the free run.

    • cites [39] Ringseis, 2019, Hepatic transcript profiling in early-lactation dairy cows fed rumen-protected niacin during the transition from late pregnancy to lactation, J. Dairy Sci. source

      Not checked in the free run.

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

    Study design and conductnone given

    Not normally distributed data were log-transformed prior to two-way ANOVA.

    “Not normally distributed data were log-transformed prior to two-way ANOVA.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    The basal diet was sufficient to meet the requirements of the rat for maintenance according to the National Research Council.

    “All groups received the same basal diet (Table 4), which was sufficient to meet the requirements of the rat for maintenance according to the National Research Council (NRC) [31]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] National Research Council (1995). Nutrient Requirements of Laboratory Animals, National Academy Press. [4th ed.].

      Not checked in the free run.

35 of 35

  • The paper relies on this without support.

    The obese Zucker rat had not previously been used to investigate the effects of ecdysterone.

    “the obese Zucker rat is a genetic rodent model for obesity, fatty liver, and hyperlipidaemia, which has not yet been used to investigate the effects of ecdysterone.”

  • The paper relies on this without support.

    The study had sufficient statistical power to detect a lipid-lowering effect of ecdysterone in Zucker rats.

    “hepatic and plasma triglyceride and cholesterol concentrations did not differ between rats of the same genotype fed the diets without or with ecdysterone.”

  • The paper relies on this without support.

    The obese Zucker rat is a valid model for human obesity, fatty liver, and hyperlipidaemia.

    “the obese Zucker rat is a genetic rodent model for obesity, fatty liver, and hyperlipidaemia”

  • The paper relies on this without support.

    Ecdysterone supplemented in the diet at 0.5 g/kg is absorbed and reaches the liver and skeletal muscle at bioactive concentrations.

    “supplemented (groups LE and OE) or not (groups LC and OC) with 0.5 g ecdysterone (provided from Alibaba, China) per kg diet.”

  • The paper relies on this without support.

    The 4-week supplementation period is sufficient to detect effects on hepatic transcriptome and lipid metabolism if they exist.

    “The rats of all groups had free access to their diets which were fed for 4 weeks.”

  • The paper relies on this without support.

    The microarray analysis with six samples per group and the chosen filter criteria has adequate statistical power to detect relevant changes.

    “For microarray analysis, six liver total RNA samples/group were randomly selected.”

  • The paper relies on this without support.

    The Zucker rat model is a valid model for human obesity and metabolic syndrome.

    “The experiment included 16 male, 25-week-old, homozygous (fa/fa) obese Zucker rats (Crl:ZUC-Leprfa) and 16 male, 25-week-old, heterozygous (fa/+) lean Zucker rats”
7 of 7

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?

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.

  • Conflicting identity of the 44.5-fold upregulated transcript

    Minor

    The results identify Sdc3-like as the most strongly upregulated transcript at 44.5-fold, while the discussion identifies Scd3-like at the same fold change. If these are distinct transcripts, Scd3-like should also appear among the ten most strongly upregulated transcripts, ahead of those listed with smaller fold changes.

1 of 1

Cites

Flagged first, then by citations

1 of the 41 references couldn’t be matched to a known paper.

Research works printed but not linked: 4

  1. Stopka, 1999, Effect of insect hormone, 20-hydroxyecdysone on growth and reproduction in mice, Acta Soc. Zoo. Bohem., 63, 367
  2. National Research Council (1995). Nutrient Requirements of Laboratory Animals, National Academy Press. [4th ed.].
  3. Sreejit, 2019, Isolation, quantification and chemical characterization of ecdysterone from medicinal plants of Kerala, Western Ghats, J. Pharmacogn. Phytochem., 8, 254
  4. VDLUFA (Verband Deutscher Landwirtschaftlicher Untersuchungs- und Forschungsanstalten) (2007). Die chemische Untersuchung von Futtermitteln. VDLUFA-Methodenbuch. Band III, Ergänzungslieferungen von 1983, 1988, 1992, 1997, 2004, 2006, 2007, VDLUFA-Verlag.

Cited by

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Related

Authors
Marschall, M. J. M., Ringseis, R., Gessner, D. K., Grundmann, S. M., Most, E., Wen, G., Maheshwari, G., Zorn, H., & Eder, K.
Affiliations
Justus-Liebig-Universität Gießen (DE)
Corresponding author
Ringseis, Robert
ORCID
Venue
International Journal of Molecular Sciences
Publisher
MDPI AG
Published
2021-05-15
Volume
22
Issue
10
Pages
5241
ISSN
1422-0067 (ISSN-L)
DOI
10.3390/ijms22105241
PMC
PMC8156757
OpenAlex
W3161880053
Cited by
13
References
41
Citations by year
2026: 3 · 2025: 2 · 2024: 2 · 2023: 4 · 2022: 1 · 2021: 1
FWCI
1.88
Type
journal article
Language
en
Open access
Licence
Locations
  • International Journal of Molecular Sciences (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Publikationsdatenbank der Fraunhofer-Gesellschaft (Fraunhofer-Gesellschaft) (submittedVersion, other-oa)
  • Fraunhofer-Publica (Fraunhofer-Gesellschaft) (submittedVersion)
  • Universitätsbibliothek Gießen (cc-by)
Funders
Deutscher Akademischer Austauschdienst (57243780)
Topics
  • Health Sciences › Medicine › Surgery › Cholesterol and Lipid Metabolism
  • Health Sciences › Medicine › Epidemiology › Liver Disease Diagnosis and Treatment
  • Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Lipid metabolism and disorders
Keywords
20-hydroxyecdysone, liver transcriptome, lipid metabolism, obesity, Zucker rats, triglycerides, cholesterol, hepatic lipid synthesis, dietary supplementation, hepatic steatosis, plasma lipids
MeSH terms
Animals, Ecdysterone, Gene Expression Regulation, Genotype, Lipids, Liver, Male, Obesity, Organ Size, Rats, Zucker, Reproducibility of Results, Fructosamine, Dietary Supplements, Gene Expression Profiling, Lipid Metabolism, Rats

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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