ارزیابی اثر ضد لیشمانیایی عصاره سیاه تخمه روی پروماستیگوت های لیشمانیا ماژور به روش MTT

Title

EVALUATION OF ANTI LEISHMANIAL EFFECT OF AGROSTEMMA GITHAGO ON LEISHMANIA MAJOR PROMASTIGOTE BY MTT ASSAY

English Abstract

Background Resistance to the first-line anti leishmanial drugs, pentavalent antimonials, are increasing around the world. Second-line drugs such as Amphotricine B and Miltefosine have also side effects and expensive for patients. According to the anti-cancer and cytotoxic effects of Agrostemma githago extract, this study was conducted to evaluate the anti leishmanial effect of Agrostemma githago in comparison Glucantime on Leishmania major promastigotes by MTT assay. Methods A number of 2.5×106 Leishmania major promastigotes in BHI medium in stationary phase were added to each well of the 96 well culture plate. Agrostemma githago extract was added to the wells at different concentrations (1, 2, 3, 4, 6, 8, 12, 16 mg/ml). Glucantime was used as a control group. Cells were incubated for 48 hours in 24°C. After centrifuged for 10 minutes in 4°C, supernatant were discarded and 50 µl of MTT were added. After 3 hours in 24°C and centrifuging in 4°C, the supernatant was removed and 100 µl of DMSO was added. After 15 minutes the plate was read by ELISA reader in 570 nm. Furthermore, Trypan blue assaywas performed to evaluate agrostemma extract effect on Leishmania major promastigotes. Results MTT results displayed that increasing concentrations of Agrostemma extract could significantly reduce cell viability of Leishmania major promastigots compared to the control group (p<0.05). Also, inhibitory effect of Agrostemma extract was dose dependent. IC50 of the Agrostemma and Glucantime were calculated 0.365 and 71.01 mg/ml, respectively. Moreover, trypan blue assay showed that increasing concentrations of the extract reduced number of live Leishmania major promastigots. Conclusions Aqueous extract of Agrostemma githago has stronger inhibitory effect than Glucantim on Leishmania major promastigots. Although more expended studies are needed to evaluate its effect in “in vivo” conditions and on host cells

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