Examination of the anabolic activity and mechanisms of action of the combination of Diosgenin and Ecdysterone in C2C12 myotubes
Open accessAbstract
Plant steroids such as ecdysterone (ECDY) or diosgenin (DIO) have been associated with anabolic and performance-enhancing effects for years. However, the molecular mechanisms have not yet been extensively studied in skeletal muscle cells. Consequently, the anabolic activity and associated molecular mechanisms of ECDY and DIO alone and in combination were investigated in C2C12 myotubes. Dose-dependent effects of both compounds on myotube diameter, mRNA expression of IGF-1 and PI3KR1 as well as expression of myosin heavy chain (MHC) proteins were analyzed in differentiated C2C12 cells. In addition, the binding affinities to androgen and estrogen receptors were analyzed. Treatment with ECDY and DIO significantly induced hypertrophy of C2C12 myotubes. Partially additive effects were observed. This is supported by the mRNA expression of IGF-1 and PI3KR1 as well as in the expression of MHC. However, no clear statement can be made regarding which combination has the strongest additive effects. Besides the results suggest that, in contrast to ECDY, DIO has antiandrogenic effects and bind on AR. Consequently, it indicate that two different mechanisms of action are activated in ECDY and DIO combinations. However, this must be confirmed in further cell cultures studies and human interventions concerning anti-doping regulations.
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In C2C12 cells, three of five combinations of 20-hydroxyecdysone and diosgenin showed significantly higher hypertrophic effects than the individual substances.
“Kostov et al. [21] investigated the combined effect of 20E and DSG in C2C12 cells at different dosages and showed significantly higher hypertrophic effects in three of the five combinations compared to the individual substances [21].”
From How reliable is the labeling of a commercial phytosteroid product? A 12-week randomized double-blind training study (2025) · cites this paper as [21]
The C2C12 cell line-based assay is well-established and has been used in numerous other studies.
“a well-established C2C12 cell line-based assay, that has been used in numerous other studies [7,21,66–68].”
From How reliable is the labeling of a commercial phytosteroid product? A 12-week randomized double-blind training study (2025) · cites this paper as [21]
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References 32 · Cited by 7
- Authors
- Kostov, T., Diel, P., & Isenmann, E.
- Affiliations
- German Sport University Cologne (DE)
- Corresponding author
- Isenmann, E.
- Venue
- Toxicology Letters
- Publisher
- Elsevier BV
- Published
- 2024-11
- Volume
- 401
- Pages
- 181-189
- ISSN
- 0378-4274 (ISSN-L)
- OpenAlex
- W4403286338
- Cited by
- 7
- References
- 32
- Citations by year
- 2026: 5 · 2025: 2
- FWCI
- 5.00
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Toxicology Letters (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- World Anti-Doping Agency (21C19EI)
- Topics
- Social Sciences › Sociology and Political Science › Doping in Sports
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Muscle Physiology and Disorders
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Hormonal and reproductive studies
- Keywords
- diosgenin, 20-hydroxyecdysone, plant steroids, muscle hypertrophy, C2C12 myotubes, insulin-like growth factor I, myosin heavy chain, androgen receptor, estrogen receptor, antiandrogenic effects, additive effects, anabolic agents, anti-doping regulations
- MeSH terms
- Animals, Cell Line, Diosgenin, Dose-Response Relationship, Drug, Drug Synergism, Ecdysterone, Insulin-Like Growth Factor I, Receptors, Androgen, Receptors, Estrogen, RNA, Messenger, Muscle Fibers, Skeletal, Myosin Heavy Chains, Anabolic Agents, Mice
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