Selective inhibition of Leishmania donovani by active extracts of wild mushrooms used by the tribal population of India: An in vitro exploration for new leads against parasitic protozoans

Successful Therapy of Murine Visceral Leishmaniasis with Astrakurkurone, a Triterpene Isolated from the Mushroom Astraeus hygrometricus, Involves the Induction of Protective Cell-Mediated Immunity and TLR9

“…We initiated our work with the active extracts of Astraeus hygrometricus (Pers.) Morgan and reported the differential antileishmanial effect against L. donovani promastigotes and intracellular amastigotes in vitro (15). In continuation with our line of investigation on antileishmanial leads, we also claimed simultaneously the isolation and structure elucidation of a novel triterpene molecule, astrakurkurone, from an active fraction of A. hygrometricus, with a significant effect against promastigotes (14).…”

“…To date, a very limited approach has been made to establish the mushrooms or mushroom-derived metabolites as therapeutic or immunostimulatory agents against Leishmania infection. As few as only seven reports, including three from our group, have been documented up to 2015 regarding the antileishmanial effect of extracts or active constituents of wild mushrooms (10)(11)(12)(13)(14)(15)(16). The first breakthrough was made by Jin and Zjawiony in 2006, when they isolated a novel compound, 5-heptadeca-8=Z,11=Z,16-trienylresorcinol, from a polypore mushroom, Merulius incarnatus Schweinitz 1822 (Corticiaceae), which inhibited the Leishmania growth in vitro (50% inhibitory concentration [IC 50 ] of 3.6 g/ml) with no toxicity on Vero cells up to 25 g/ml (10).…”

Protective effect of Croton caudatus Geisel leaf extract against experimental visceral leishmaniasis induces proinflammatory cytokines in vitro and in vivo

“…The total protein content of treated promastigotes was assessed by staining with FITC, an acidic dye that binds covalently to the positively charged groups of proteins as described earlier (Kamau et al, 2000;Mallick et al, 2014). Briefly, 4 × 10 6 /mL log phase promastigotes were cultured without or with 50% inhibitory concentration of JDHex and continued for 24 h and 48 h. After washing with PBS, cells were treated with 4 μg/mL DNase (Type 1, SigmaAldrich) at 37°C for 10 min.…”

“…The chromatin condensation and nuclear damage in JDHextreated promastigotes have been detected under confocal microscope after staining with DAPI as described earlier (Mallick et al, 2014). Control and treated promastigotes were washed with 1X cold PBS (×2), allowed to adhere on 0.01% poly-l-lysine coated 8-chambered slide (15 min) and fixed with 4% paraformaldehyde in 1X phosphate buffer (pH 7.2) for 30 min at room temperature.…”

“…Neutral lipid and polar lipid droplets were assessed as described earlier (Greenspan et al, 1985;Mallick et al, 2014). L. donovani promastigotes (1 × 10 6 cells/mL) in log phase were incubated at 22°C in complete M199 medium, and incubated with 10 μg/mL of JDHex (IC50 against promastigotes) in triplicate for 24 h and 48 h. Cells were centrifuged and resuspended in PBS buffer.…”