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Aim To investigate the protective effect of orcinol glucoside on dexamethasone(DEX)-induced osteoblast injury and its mechanism. Methods Primary osteoblasts were extracted from calvaria of neonatal mice and cultured in medium with DEX(1 μmol•L
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ObjectiveTo predict the therapeutic targets and related signaling pathways of orcinol glucoside （OG） in the treatment of osteoporosis by network pharmacology， and further clarify its mechanisms based on molecular docking and in vitro cell model. MethodThe pharmacological targets of OG were obtained from Similarity ensemble approach （SEA） and SwissTargetPrediction， and the targets related to osteoporosis from DisGeNET and GeneCards. The cross-analysis was conducted to screen the common targets between OG and osteoporosis. STRING was used to construct the protein-protein interaction （PPI） network， followed by topology analysis using CytoNCA plug-in of Cytoscape 3.7.2 to screen out the core targets. The obtained common targets were subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） analysis by g：Profiler. AutoDock Vina was utilized for molecular docking， and the in vitro cell experiments were then carried out for verifying the mechanism of OG in treating osteoporosis. ResultA total of 73 targets related to OG and osteoporosis were harvested，among which 14 were proved to be key targets by topological analysis. GO and KEGG functional enrichment analysis yielded 259 cell biological processes， mainly involving organonitrogen compound metabolic process， cell population proliferation， protein metabolic process， regulation of response to stress， and response to chemicals. Its mechanism of action might be related to advanced glycation end-product （AGE）-AGE receptor （RAGE） signaling pathway， interleukin-17 （IL-17） signaling pathway， and phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. Molecular docking indicated that the binding energies of OG to Cyclin D1 （CCND1） and cyclin-dependent kinase 4 （CDK4） were the lowest and similar. The results of flow cytometry showed that compared with the normal group， OG group exhibited decreased proportion of cells in G0/G1 phase （P<0.01） and decreased proportion of cells in S phase （P<0.01）. As demonstrated by Western blot， compared with the normal group， OG up-regulated the protein expression levels of Cyclin D1 and CDK4 （P<0.05， P<0.01）. ConclusionOG alleviates osteoporosis via multiple targets and multiple pathways. It may exert the therapeutic effects by increasing Cyclin D1 and CDK4 protein expression to change cell cycle and promote cell proliferation.
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Objective: To study the secondary metabolites of Eurotium cristatum from Fu Brick Tea and their biological activities. Methods: The compounds were isolated by various column chromatographies on normal phase silica gel, reversed silica gel, Sephadex LH-20, preparative TLC, and recrystallization. The structures were identified by the extensive analysis on their spectroscopic data. The inhibitory effects of compounds 1-9 against SF-268, MCF-7, and NCI-H460 cell lines were tested in vitro by SRB method. The inhibitory effects of compounds 2-9 on Staphyloccocus aureus, Escherichia coli, Salmonella enterica subsp. enterica, Shigella dysenteriae, and Proteus vulgaris were tested by MTT method. Results: Nine compounds were isolated from the extract of liquid fermentation broth of E. cristatum in Fu Brick Tea and identified as physcion (1), 1, 5-dihydroxy-3-methoxy-7-methyl-anthracene-9,10-dione (2), flavoglaucin (3), 2-(2', 3-epoxy-1', 3'-heptadienyl)-6-hydroxy-5-(3-methyl-2-butenyl) benzaldehyde (4), 2-(2', 3-epoxy-1', 3', 5'-heptatrienyl)-6-hydroxy-5-(3-methyl-2-butenyl) benzaldehyde (5), isodihydroauroglaucin (6), tyrosol (7), p-hydroxybenzoic acid (8), and orcinol (9). The results of the bioactivity test showed that compounds 3-6 displayed the good cytotoxic acitivities against the three tumor cell lines, and all the compounds exhibited the strong inhibitory activities against P. vulgaris except compounds 1 and 6. Besides, compound 9 also showed the significant inhibitory effects against S. aureus and E. coli. Conclusion: The benzaldehyde compounds are considered as the main metabolites of E. cristatum, compounds 7 and 9 are reported from the species of genus Eurotium Link: Fr. for the first time and compounds 2, 3, 5, 6, and 8 are firstly isolated from the fungus.