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This study aimed to compare the effects of 20E and CAL on metabolic, muscular, and endocrine parameters in normal and obese male rats, with a particular focus on androgen balance. Methods: Male rats were treated with 20E or CAL under normal (ND) or high-fat, high-sugar dietary (HFHSD) conditions for 12 weeks. Body weight, food intake, skeletal and androgen-sensitive muscle mass, and testicular weight were measured. Testicular expression of androgen receptor (Ar) and aromatase (Cyp19a1) mRNA was assessed by RT-PCR. ELISA was used to determine the plasma corticosterone, testosterone and ER\u03b2 level in testes. Results: 20E and CAL prevented HFHSD-induced weight gain and skeletal muscle atrophy. CAL uniquely preserved testicular and levator ani muscle mass in obese rats. CAL increased the expression of Cyp19a1 and ER\u03b2 in testicles. Decreased Ar mRNA expression was regulated in 20E- and CAL-treated obese animals. While 20E treatment significantly reduced plasma testosterone levels in the normal diet group compared to controls, both 20E and CAL interventions elicited significant reductions in the obese group relative to both the ND and HFHSD groups. HFHSD-induced increase in plasma corticosterone levels was normalized by 20E or CAL treatment. Conclusions: 20E and CAL exhibit beneficial metabolic and anabolic effects by preventing HFHSD-induced obesity and consequential muscle atrophy. CAL counteracts obesity-induced testicular atrophy in terms of tissue mass. Based on our results, we hypothesized that CAL enhances testicular aromatase levels, which may lead to increased compensatory androgen receptor mRNA expression and increased ER\u03b2 levels. These complex, not yet fully understood results underscore the need for caution in the use of phytoecdysteroids as dietary supplements.<\/jats:p>","DOI":"10.3390\/nu18122023","type":"journal-article","created":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T06:13:37Z","timestamp":1782195217000},"page":"2023","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Prevention of Diet-Induced Obesity by Phytoecdysteroids 20-Hydroxyecdysone and Calonysterone\u2014Unexpected Modulation of Androgen Balance in Normal and Obese Rats"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8306-6053","authenticated-orcid":false,"given":"Alaa AM","family":"Osman","sequence":"first","affiliation":[{"name":"Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, E\u00f6tv\u00f6s u. 6, H-6720 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D\u00e1vid","family":"Laczk\u00f3","sequence":"additional","affiliation":[{"name":"Institute of Pharmacognosy, Faculty of Pharmacy, University of Szeged, E\u00f6tv\u00f6s u. 6, H-6720 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00e1t\u00e9","family":"V\u00e1gv\u00f6lgyi","sequence":"additional","affiliation":[{"name":"Institute of Pharmacognosy, Faculty of Pharmacy, University of Szeged, E\u00f6tv\u00f6s u. 6, H-6720 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-4167-4653","authenticated-orcid":false,"given":"No\u00e9mi","family":"T\u00f3th","sequence":"additional","affiliation":[{"name":"Institute of Pharmacognosy, Faculty of Pharmacy, University of Szeged, E\u00f6tv\u00f6s u. 6, H-6720 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2746-5372","authenticated-orcid":false,"given":"Kata Kira","family":"Kem\u00e9ny","sequence":"additional","affiliation":[{"name":"Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, E\u00f6tv\u00f6s u. 6, H-6720 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-1408-3286","authenticated-orcid":false,"given":"P\u00e9ter","family":"Szatm\u00e1ri","sequence":"additional","affiliation":[{"name":"Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, E\u00f6tv\u00f6s u. 6, H-6720 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrienn","family":"Seres-Bokor","sequence":"additional","affiliation":[{"name":"Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, E\u00f6tv\u00f6s u. 6, H-6720 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0074-3472","authenticated-orcid":false,"given":"Attila","family":"Hunyadi","sequence":"additional","affiliation":[{"name":"Institute of Pharmacognosy, Faculty of Pharmacy, University of Szeged, E\u00f6tv\u00f6s u. 6, H-6720 Szeged, Hungary"},{"name":"HUN-REN-SZTE Biologically Active Natural Products Research Group, E\u00f6tv\u00f6s u. 6, H-6720 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eszter","family":"Ducza","sequence":"additional","affiliation":[{"name":"Department of Pharmacodynamics and Biopharmacy, Faculty of Pharmacy, University of Szeged, E\u00f6tv\u00f6s u. 6, H-6720 Szeged, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,21]]},"reference":[{"key":"ref_1","unstructured":"(2026, June 18). 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