Talaromyces purpurogenus from a marine-polluted environment inhibits pro-inflammatory markers in LPS-stimulated RAW 264.7 cells and in carrageenan-induced paw edema

Abstract

ObjectivesTalaromyces purpurogenus is a fungus species widely found in the marine environment. When in a hostile environment, it produces a series of secondary metabolites with important biotechnological potential, such as meroterpenoids. In our previous report, we demonstrated that the culture broth ethyl acetate extract from T. purpurogenus reduced the cellular infiltration of lymphoid organs and circulating levels of TNF-α. To elucidate this anti-inflammatory effect, this work aimed to study T. purpurogenus isolated from a marine-polluted environment regarding the chemical composition of its culture broth and mycelial ethyl acetate extract (CBEAE and MEAE respectively), as well as its action on pro-inflammatory markers in LPS-stimulated RAW 264.7 cells and carrageenan-induced mice paw edema. MethodsChemical characterization was performed by HPLC-DAD-UV. Cytotoxicity in vitro and endotoxin quantification in extracts were preformed prior the experiments. In vitro inhibitory effect of T. purpurogenus extracts were evaluated by cytokines and nitrite quantification in cell supernatant of RAW 264.7 cells. The effect of T. purpurogenus in murine acute inflammation model of carrageenan-induced paw edema was verified by thickness kinetic and histology evaluation. ResultsThe CBEAE demonstrated a richer chemical composition than MEAE, presenting nine meroterpenoid compounds, while MEAE displayed only two meroterpenoid and one phenolic compound. Cytotoxicity in RAW 264.7 macrophages was not observed at any concentration treatment for both extracts. Endotoxin quantification also demonstrated absence of endotoxins in both extracts. At 500 μg/mL, the CBEAE and MEAE extracts inhibited nitrite (p = 0.0450 and p = 0.0497), TNF-α (p = 0.0076 and p = 0.0001) and IL-1β (p = 0.0042 and p = 0.0031), but only CBEAE inhibited IL-6 levels in LPS-stimulated cells (p = 0.0234). CBEAE, but not MEAE, also inhibited nitrite (p = 0.0353) and IL-1β (p = 0.0386) in non-stimulated cells. In addition, CBEAE at 250 and 500 mg/kg inhibited edema thickness and mast cells (p = 0.0303 and p = 0.0130) in λ-carrageenan-induced mice paw edema. ConclusionsTalaromyces purpurogenus isolated from marine-polluted environment inhibited pro-inflammatory markers in LPS-stimulated RAW 264.7 cells and in murine model of paw edema. The promising anti-inflammatory activity in vitro and in vivo, especially the culture broth ethyl acetate extract, demonstrates its potential for further studies to elucidate its mechanism of action.