Bioactive compounds from Psilocybe cubensis mycelial cultures with transient anxiolytic effects in mice.

The growing percentage of people suffering from drug-resistant depression increases interest in alternative therapies, particularly the usage of psychedelics such as psilocybin. The main source of psilocybin is the Psilocybe cubensis species. Due to the potential therapeutic benefits of psilocybin and the legal restrictions on its possession and use in the form of fungal fruiting bodies, this research work documents an attempt to obtain in vitro P. cubensis mycelium in which psilocybin and other biologically active substances acting on the central nervous system would be present. It was hypothesized that chronic microdosing with whole in vitro - cultured P. cubensis mycelium, containing psilocybin together with other neuroactive secondary metabolites, could exert anti-anxiety and antidepressant effects through their combined action. For this purpose, the anti-anxiety and antidepressant activity of mycelium microdosing in male C57BL/6J mice was investigated. The tail suspension test (TST), novelty suppressed feeding test (NSFT), sucrose preference test (SPT), locomotor activity (LA), and female urine sniffing test (FUST) were used to examine animal behavior. The chemical analysis was performed using high-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC) method. Analysis of the studied extracts showed that psilocybin was present only in the mycelium of the Cambodian strain, at a concentration of 20.78 mg per 100 g dry weight. The experiment showed that mycelium supplementation significantly reduced anxious behavior in mice on day 22 but did not affect locomotor activity, depressive, anxiety-related, or anhedonic behaviors at later stages of the experimental protocol. Although the results suggest the potential of P. cubensis mycelial cultures in anxiety prevention, further studies using higher doses or alternative models are needed to confirm and extend these findings.