In vitro antileishmanial activity of fisetin flavonoid via inhibition of glutathione biosynthesis and arginase activity in Leishmania infantum

Abstract

With the increasing emergence of drug resistant Leishmania sp. in recent years, combination therapy has been considered as a useful way to treat and control of Leishmaniasis. The present study was designed to evaluate the antileishmanial effects of the fisetin alone and combination of fisetin plus Meglumine antimoniate (Fi-MA) against Leishmania infantum. The IC50 values for fisetin were obtained 0.283 and 0.102 μM against promastigotes and amastigote forms, respectively. Meglumine antimoniate (MA, Glucantime) as control drug also revealed IC50 values of 0.247 and 0.105 μM for promastigotes and amastigotes of L. infantum, respectively. In order to determine the mode of action of fisetin and Meglumine antimoniate (MA, Glucantime), the activities of arginase (ARG), catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) were measured. Moreover, intracellular glutathione (GSH) and nitric oxide (NO) levels in L. infantum-infected macrophages and L. infantum promastigotes which were treated with IC50 concentrations of fisetin, MA and Fi-MA were investigated. Our results showed that MA decreased CAT and SOD activity and increased NO levels in L. infantum-infected macrophages. In promastigotes, MA inhibited parasite SOD activity and reduced parasite NO production. The decreased levels of most of the antioxidant enzymes, accompanying by the raised level of NO in treated macrophages with MA, were observed to regain their normal profiles due to Fi-MA treatment. Furthermore, fisetin could prevent the growth of promastigotes by inhibition of ARG activity and reduction of GSH levels and NO production. In conclusion, these findings showed that fisetin improves MA side effects.

Flavonoids as efficient scaffolds: Recent trends for malaria, leishmaniasis, Chagas disease, and dengue

“…Some other antileishmanial compounds with low IC 50 , that is, compounds 7 (IC 50 : 0.1390 μM); 103 (IC 50 : 0.0513 μM); and 104 (IC 50 : 0.0446 μM; Gontijo et al, ), were also nominated as hit compounds. In visceral leishmaniasis, fisetin ( 37 ; IC 50 : 0.102 μM; SI: 239.01, Raw 264.7 cells; Adinehbeigi et al, ) and ageconyflavone C ( 114 ; IC 50 : 3.56 μg/ml; SI: >25, L6 cells) can be considered as hit compounds. In antitrypanosomal studies, potential hit compounds included astragalin heptaacetate ( 122 ; 0.8 μM; SI: 205, Vero cells; Marín et al, ); 5‐desmethylsinensetin ( 85 ; IC 50 : 0.2 μg/ml; SI: >500); and eupatorin ( 86 ; IC 50 : 0.4 μg/ml; SI: >500; Beer et al, ).…”

“…From the database searches, 830 (“malaria and flavonoids”: 540; “Chagas disease and flavonoids”: 60; “leishmaniasis and flavonoids”: 124; “dengue and flavonoids”: 106) potentially relevant records were identified, from which 737 were excluded after screening the titles or abstracts. The full reports of 93 articles were acquired: 83 studies related to natural flavonoids (Adinehbeigi, Razi Jalali, Shahriari, & Bahrami, ; Allard et al, ; Anusuya & Gromiha, ; Anusuya & Gromiha, ; Assis et al, ; Azebaze et al, ; Beer et al, ; Cabanillas et al, ; Castro, Barrios, Chinchilla, & Guerrero, ; Cheema et al, ; Cornelio et al, ; Coulerie et al, ; Cunha‐Rodrigues et al, ; Da Rocha et al, ; Da Silva, Maquiaveli, & Magalhães, ; De Monbrison et al, ; De Sousa et al, ; De Sousa et al, ; Dos Reis, Manjolin, Maquiaveli, Santos‐Filho, & da Silva, ; Ezenyi et al, ; Fonseca‐Silva, Inacio, Canto‐Cavalheiro, & Almeida‐Amaral, ; Fonseca‐Silva, Inacio, Canto‐Cavalheiro, Menna‐Barreto, & Almeida‐Amaral, ; Frabasile et al, ; Freitas et al, ; Ganesh et al, ; Gontijo et al, ; Grael, Albuquerque, & Lopes, ; Grecco et al, ; Güida et al, ; Heh et al, ; Houël et al, ; Ichino et al, ; Ijaz, Ahmad, Ahmad, ul Haq, & Wang, ; Inacio, Gervazoni, Canto‐Cavalheiro, & Almeida‐Amaral, ; Ismail & Jusoh, ; Jasmeen et al, ; Jin et al, ; Kapingu et al, ; Kiat et al, ; Kimmel et al, ; Konziase, ; Kraft et al, ; Kraft et al, ; Lage et al, ; Lage et al, ; Lehane & Saliba, ; Leite et al, ; Lu et al, ; Manjolin, dos Reis, Maquiaveli, Santos‐Filho, & da Silva, ; Marín et al, ; Marín et al, ; Mittra et al, ; Moghaddam et al, ; Montenegro, Gonzalez, Ortega‐Barria, & Luis Cubi...…”

“…Moreover, compounds 112 and 113 were not selective for the Leishmania parasite as compared with L6 cells. After 48 hr of incubation, fisetin inhibited promastigote and intramacrophage amastigotes of L. infantum , with IC 50 values of 0.283 μM (SI: 86.14; Raw 264.7 cells) and 0.102 μM (SI: 239.01; Raw 264.7 cells), respectively (Adinehbeigi et al, ).…”

Efficacy of Topical Treatment with (−)-Epigallocatechin Gallate, A Green Tea Catechin, in Mice with Cutaneous Leishmaniasis

“…The relative amount of formazan produced by viable macrophages was determined using a spectrophotometer at 570 nm. The percentage of viable cells was calculated according to the formula: OD treated culture-OD blank x 100/OD control-OD blank as was described previously [19]. The CC 50 was calculated for each compound as the concentration of compound that reduces cell growth by 50%.…”

Leishmania infantum arginase: biochemical characterization and inhibition by naturally occurring phenolic substances

“…To the best of our knowledge, there is a single report on Leishmania infantum arginase. In this specific study, enzyme inhibition assay was performed with cellular extracts and did not employ the purified enzyme 20 . Here, we biochemically characterised recombinant arginase from L. infantum (herein referred as ARGLi) and evaluated its inhibition by a panel of fourteen naturally occurring phenolic substances.…”