Evaluation of the Paraquat Effect on Leishmania major Promastigotes and HUVECs Viability under in vitro Condition

Abstract

Background & objectives: Pentavalent antimonials are the first-line drugs for treatment of leishmaniasis, which have multiple side effects such as drug toxicity. Moreover, parasite resistance to these drugs is rising around the world. Second-line drugs, including Amphotericin B and pantamidine have also side effects and expensive for patients. According to the cytotoxic effects of paraquat, this study was conducted to evaluate the effect of paraquat on Leishmania major promastigotes and HUVECs viability. Methods: A number of 2.5×106 of Leishmania major promastigotes were treated in each well of 96 well plates with different concentrations of paraquat. Cells were incubated for 48 hours in 24 °C. MTT test was performed for evaluating paraquat impact on promastigotes. The absorbance was measured using a microplate reader at 570 nm. The trypan blue staining assay was performed to evaluate the number of viable Leishmania major promastigotes following paraquat treatment. Furthermore, the effect of paraquat concentrations on HUVECs viability was evaluated under the cell culture condition. Results: The results of the MTT test showed that increasing concentrations of paraquat could significantly reduce the viability and the number of Leishmania major promastigotes in comparison to control group (p<0.05). In this study, the IC50 for Leishmania major promastigotes was calculated as 272.46 µg/ml. Trypan blue results were in line with the finding of MTT assay. Moreover, we found that HUVECs were susceptible to paraquat (IC50=188.99 µg/ml). Conclusion: Paraquat has a strong inhibitory effect on Leishmania major promastigotes and human endothelial cells. Although more comprehensive studies on the effects of the topical use of paraquat on Leishmania major lesions in animal model and its side effects are necessary.

Keywords

• Leishmania major
• Paraquat
• MTT Assay