The Influence of Seasonality on Secondary Metabolite Profiles and Neuroprotective Activities of Moss Hypnum cupressiforme Extracts: In Vitro and In Silico Study

Abstract

Numerous representatives of mosses, including Hypnum cupressiforme, have been used toalleviate different inflammation-related conditions. However, the mode of action underlying thisanti-inflammatory potential has been poorly understood. Moreover, the influence of seasonalityon the chemical composition and biological activity of mosses is generally overlooked. This studyaimed to investigate the influence of seasonal changes (spring, summer, and autumn) on secondarymetabolite composition and biological activities of ethyl acetate H. cupressiforme extracts. Antioxidantactivity was measured usingβ-carotene bleaching assay, while MTT, NBT, ELISA, and Griess assayswere carried out to explore the anti-neuroinflammatory and neuroprotective potential of extracts.Inhibitory activities on acetylcholinesterase and tyrosinase were assessed experimentally and bydocking analysis. The highest content of secondary metabolites and antioxidant activity wereobserved in moss during the summer. Extracts inhibited the secretion of ROS, NO, TNF-α, andIL-6, alleviating the inflammatory potential of H2O2and LPS in microglial and neuronal cells.Strong inhibitory effects on acetylcholinesterase and tyrosinase were observedin vitro. Dockinganalyses revealed high-affinity interactions of secondary metabolites present in H. cupressiforme withimportant enzyme residues. Altogether, these results reveal the neuroprotective potential and thesignificance of seasonal fluctuations on secondary metabolite content and biological activities in mossH. cupressiforme