Leishmanicidal and immunomodulatory activities of the formononetin (a natural isoflavone) against Leishmania tropica

Abstract

Objective: This work aimed to examine the leishmanicidal, cellular mechanisms and cytotoxicity effects of formononetin (FMN), a natural isoflavone, against Leishmania tropica. We used the MTT assay to determine the leishmanicidal effects of FMN against promastigotes and its cytotoxicity effects on J774-A1 macrophage cells. The Griess reaction assay and quantitative real-time PCR were used to determine the nitric oxide (NO) and the mRNA expression levels of IFN-γ and iNOS in infected J774-A1 macrophage cells.

Results: FMN significantly (P < 0.001) decreased the viability and number of promastigotes and amastigotes forms. The 50% inhibitory concentrations value for FMN and glucantime was 9.3 and 14.3 µM for promastigote and amastigote, respectively. We found that the macrophages exposed with FMN especially at concentrations of 1/2 IC₅₀ and IC₅₀ significantly activated the NO release and the mRNA expression levels of IFN-γ, iNOS. The findings of the current research showed the favorable antileishmanial effects formononetin, a natural isoflavone, against various stages of L. tropica through inhibition of infectivity rate of macrophage cells and triggering the NO production and cellular immunity. However, supplementary works are essential to evaluate the ability and safety of FMN in animal model before use in the clinical phase.

Keywords: Amastigote; Infectivity; Leishmaniasis; Nitric oxide; Promastigote.

Fig. 1

Inhibitory effects of formononetin (FMN), meglumin antimoniate (MA), and amphotericin B (AmB) against Leishmania tropica promastigote (A), amastigote (B), and macrophage cells (C). Mean ± standard deviation. (n = 3)

Fig. 2

mRNA expression of genes (iNOS and IFNγ) in infected macrophages with Leishmania tropica promastigotes treated with formononetin (FMN) in comparison with the control group. mean ± SD (n = 3). * p < 0.001