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RNAseq analysis on liver from pigs treated with testosterone and nandrolone esters: identification and field validation of transcriptional biomarkers

ABSTRACT: The use of anabolic steroid hormones as growth promoters in farm animals is banned in the European Union. However, among anabolic compounds, testosterone and nandrolone are still illegally administered to fattening pigs in the form of synthetic steroid esters. To tackle steroid misuse alternative approaches based on biological methods, able to investigate perturbations induced by these compounds at level of proteins, transcripts, metabolites is becoming recommendable. The aim of this work was to characterize by RNAseq coding mRNAs perturbations related to illicit administration of testosterone and nandrolone esters in fattening pigs. Twenty-seven clinically healthy 90-day-old pigs were randomly assigned to test (18 animals) and control (9 animals) groups. Nine of the eighteen animals from the test group were treated with an i.m. injections of 4 mg/kg b.w. of Sustanon (250 mg/ml). The other nine animals of the test group undergo i.m. injections of 5 mg/kg b.w. of Myodine (25 mg/ml). The animals were injected on day 4 after the start of the experiment and were euthanized on day 114 of the experiment. Liver samples were collected and stored in RNAlater solution for gene expression studies.

ORGANISM(S): Sus scrofa

PROVIDER: GSE233177 | GEO | 2023/11/29

REPOSITORIES: GEO

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Project description:Glucocorticoids are a well recognized and common cause of muscle atrophy. Glucocorticoid-induced atrophy can be prevented by testosterone, but the molecular mechanisms underlying such protection have not been described. Thus, the global effects of testosterone on dexamethasone-induced changes in gene expression were evaluated in rat gastrocnemius muscle using Affymetrix 230_2 DNA microarrays. Gene expression was analyzed after 7 days administration of dexamethasone, dexamethasone plus testosterone, or vehicle. Effects of these agents on weights of gastrocnemius muscles from these animals has been reported (1. Zhao W, Pan J, Zhao Z, Wu Y, Bauman WA, and Cardozo CP. Testosterone protects against dexamethasone-induced muscle atrophy, protein degradation and MAFbx upregulation. J Steroid Biochem Mol Biol 110: 125-129, 2008.) Dexamethasone changed expression of 876 probe sets by at least 2-fold, of which 474 probe sets were changed by at least two fold in the opposite direction in the dexamethasone plus testosterone group (genes in opposition). Major biological themes represented by genes in opposition included IGF-1 signaling, protein synthesis, myogenesis and muscle development, and ubiquitin conjugases and ligases. Testosterone blocked increased expression of DDIT4 and eIF4EBP1, FOXO1 and of the p85 regulatory subunit of the IGF-1 receptor, while preventing decreased expression of IRS-1. Testosterone blocked decreased expression of LXR and suppressed upregulation of C/EBP beta and delta. Testosterone prevented increase expression of Cdkn1A (p21) and decrease expression of cyclins B and D, as well as many other changes that would be expected to reduce cell cycle progression. Testosterone prevented increased expression of muscle development factors Csrp3 and Mbn1 and blocked reduced expression of Wnt4. These data suggest that testosterone blocks multiple changes in gene expression that, collectively, would otherwise downregulate molecular signals that promote protein synthesis and muscle hypertrophy and that stimulate muscle protein catabolism. Experiment Overall Design: Rats (250 g males) were administered dexamethasone (0.7 mg/kg/day) alone or with testosterone (20 mg/kg/day) dissolved in vehicle (propylene glycol) by continuous infusion (Alzet pumps) for 7 days. Control animals were administered vehicle only. Gene expression was compared among the 3 groups at 7 days using one chip per animal, 3 animals per group.

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RNAseq analysis on liver from pigs treated with testosterone and nandrolone esters: identification and field validation of transcriptional biomarkers

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