Antileishmanial Activity of Sodium Stibogluconate

RAW 264.7 cells (~ 5.0 × 10⁵ cells/per well) were plated on round glass coverslips in 24-well plates and allowed to adhere to the slides for 24 h at 37 °C, 5% CO₂ with RPMI-1640 medium (Sigma-Aldrich) supplemented with 10% FBS (Gibco). Stationary phase promastigotes (~ 5.0 × 10⁶ cells/per well) of HCZ isolate, DD8 and 9044 strains were added into wells, and incubated at 37 °C, 5% CO₂ for 6 h. Next, the non-infected promastigotes were removed, and the infected macrophages were incubated at 37 °C in 5% CO₂ with fresh medium without drugs for 24 h. The medium was then removed, and fresh culture medium supplemented with 10 μM SSG (cf. [33 (link)]) were added into wells for 2 days, with PBS used as control. At 24 and 48 h, the glass coverslips were fixed in methanol (Solarbio, Beijing, China) and stained with Wright’s stain (Solarbio) to calculate the number of intracellular amastigotes under light microscopy by counting 200 macrophages per slide. Inhibition rate (%) = 100% − [(No. of amastigotes in treated sample/ No. of amastigotes in control) × 100%].