In vitro Anti Leishmanial Effect of Agrostemma githago Extract on Leishmania Major Promastigotes by Cell Count and MTT Assay

Abstract

Background and purpose: Treatment of leishmaniasis is getting complicated due to multiple side effects and drug resistance to first-line drugs. Agrostemma githago is a plant with anti-cancer and cytocidal effects, so, this study was conducted to evaluate the anti leishmanial effect of its extract on Leishmania major promastigotes by MTT assay and cell count. Materials and methods: A total of 2.5×106 Leishmania major promastigotes in their stationary phase were plated to each well of the 96 well culture plates. Cells were then incubated with increasing concentrations of Agrostemma githago extract (0.05 – 2.4 mg/ml) for 48 hours at 25°C. Glucantim was used as standard control. Then, the supernatants were discarded and 50 µl of MTT were added for 3 hours. After centrifuge, the supernatants were replaced by 100 µl of DMSO. The plate was read by ELISA reader at 570 nm. Trypan blue staining was also performed to evaluate the effect of Agrostemma githago extract on Leishmania major promastigotes. Results: MTT assay showed that increasing concentrations of Agrostemma extract could significantly reduce cell viability of Leishmania major promastigots in a dose dependent manner (p<0.05). IC50 of the Agrostemma and Glucantime were 0.365 and 71.01 mg/ml, respectively. Conclusion: Aqueous extract of Agrostemma githago was found to have stronger inhibitory effect than Glucantim on Leishmania major promastigots.

Keywords

• leishmania major
• agrostemma githago
• mtt assay