## 4 Discussion
Regarding the fatty acids profile, unsaturated fatty acids are prevalent both in WSWSCO2 and the vegetal biomass. These results confirm literature data on wild strawberry seeds fatty acids, which reported unsaturated fatty acids oleic, linoleic, and linoleic acids are the most abundant compared to saturated stearic acid [34]. [PERSON] et al. [15] evaluated the fatty acids composition of wild strawberry oils obtained by cold pressing and supercritical CO2 fractions collected at different timepoints of extraction. The lowest amount of polyunsaturated-rated linoleic acid (30%) was found in oil obtained by cold pressing, while in supercritical CO2 fractions, higher values were detected, up to 43%. Furthermore, monounsaturated oleic acid was higher in oil obtained by cold pressing, while in CO2 fractions was detected the lowest amount (12%), suggesting an influence in fatty acid composition depending on the extraction method and a preservation of polyunsaturated compounds with supercritical CO2 extraction technique. Moreover, our analysis confirms the preservation of fatty acids profiles and relative abundance both in waste and extract, suggesting that supercritical CO2 extraction could be a valuable approach to recover and maintain the native fatty acid composition from plants. As the next step, the in vitro modulation of the expression of 5\(\alpha\)-reductase gene in skin fibroblasts was assessed for WSWSCO2 compared to saw palmetto extract as the positive control. WSWSCO2 extract at the concentration of 5 mg/mL is able to inhibit the expression levels of 5\(\alpha\)-reductase type 1, being 500 times less active than saw palmetto. This is somewhat expected because many studies report the efficacy of saw palmetto in 5\(\alpha\)-reductase inhibition, and this ingredient is commonly found in food supplements as well as pharmaceutical preparations for the treatment of benign prostatic hypertrophy and prostate cancer and, for this reason, was chosen as the positive control in our study [35; 36; 37; 38]. Other mechanisms of actions related to saw palmetto have also been proposed, involving anti-androgenic, anti-proliferative, and anti-inflammatory effects [39]. Saw palmetto preparations are available in many dosage forms, including hard capsules containing dried extract and soft gel containing oily extract, typically standardized in fatty acids and phytosterols [40]. Among these fatty acids, the major constituents are lauric, oleic, myristic, and linoleic acids that are reported to be effective inhibitors of 5\(\alpha\)-reductase isozymes (type 1 and 2) and involved in saw palmetto mechanisms of action [41]. [PERSON] et al. studied the binding activities of major fatty acids (lauric, oleic, palmitic, myristic, and linoleic) contained in saw palmetto extract for benign prostatic hypertrophy--pharmacologically relevant (\(\alpha\)1-adrenergic, muscarinic, 1,4-DHP calcium channel antagonist) receptors. In addition, the effects of saw palmetto and its fatty acids on 5\(\alpha\)-reductase activity in rat liver were also examined. Considering the results, linoleic acid presented the greatest receptor-binding activity and inhibitory effect on 5\(\alpha\)-reductase compared to other free fatty acids contained in saw palmetto [42]. Inhibition of 5\(\alpha\)-reductase activity was reported also for linoleic acid, and the presence of unsaturated bonds was demonstrated to enhance the inhibitory activity [43]. This may suggest an important role of polyunsaturated fatty acids for pharmacodynamic mechanisms of inhibition of 5\(\alpha\)-reductase related to the saw palmetto phytocomplex. Compared to saw palmetto, the WSWSCO\({}_{2}\) extract developed in this study contains a major amount of unsaturated fatty acids, and, for this reason, the inhibitory activity against 5\(\alpha\)-reductase was investigated.