Objective:To investigate the biochemical capacity,and in vitro inhibitory effects of hairy roots from two cultivars of Ficus carica L.(Sabz and Siah)on Leishmania major promastigotes and amastigotes.Methods:In the hai...Objective:To investigate the biochemical capacity,and in vitro inhibitory effects of hairy roots from two cultivars of Ficus carica L.(Sabz and Siah)on Leishmania major promastigotes and amastigotes.Methods:In the hairy roots,the activity of antioxidant enzymes compared to normal leaves and roots,and the presence of some phenolic compounds in comparison with fruits were investigated.The IC_(50) values of hairy roots in promastigotes was determined by tetrazolium-dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue assays.By calculating the infectivity index of peripheral blood mononuclear cells(PBMCs),the leishmanicidal activity(IC_(50) values)of hairy roots for amastigotes was estimated.The effects of hairy roots(IC_(50) values)treatment on the levels of IFN-γ and iNOS expression,intracellular reactive oxygen species,and iNOS protein expression in infected-PBMCs were determined.Results:Based on antioxidant enzyme assays and high performance liquid chromatography analysis,hairy roots exhibited high antioxidant capacity and contained high levels of phenolic compounds.According to the results of tetrazolium-dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue assays,the hairy root extracts of both cultivars showed considerable dose-dependent inhibitory effects against Leishmania major promastigotes.Depending on the concentration and exposure time,treatment of infected-PBMCs with hairy root extracts caused the generation of a significant reactive oxygen species,up-regulation of IFN-γ and iNOS genes expression,and high value of iNOS protein compared to controls.Conclusions:The findings of this study suggest that the hairy roots of Ficus carica can be considered as a promising natural source of antileishmanial agents.展开更多
文摘Objective:To investigate the biochemical capacity,and in vitro inhibitory effects of hairy roots from two cultivars of Ficus carica L.(Sabz and Siah)on Leishmania major promastigotes and amastigotes.Methods:In the hairy roots,the activity of antioxidant enzymes compared to normal leaves and roots,and the presence of some phenolic compounds in comparison with fruits were investigated.The IC_(50) values of hairy roots in promastigotes was determined by tetrazolium-dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue assays.By calculating the infectivity index of peripheral blood mononuclear cells(PBMCs),the leishmanicidal activity(IC_(50) values)of hairy roots for amastigotes was estimated.The effects of hairy roots(IC_(50) values)treatment on the levels of IFN-γ and iNOS expression,intracellular reactive oxygen species,and iNOS protein expression in infected-PBMCs were determined.Results:Based on antioxidant enzyme assays and high performance liquid chromatography analysis,hairy roots exhibited high antioxidant capacity and contained high levels of phenolic compounds.According to the results of tetrazolium-dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue assays,the hairy root extracts of both cultivars showed considerable dose-dependent inhibitory effects against Leishmania major promastigotes.Depending on the concentration and exposure time,treatment of infected-PBMCs with hairy root extracts caused the generation of a significant reactive oxygen species,up-regulation of IFN-γ and iNOS genes expression,and high value of iNOS protein compared to controls.Conclusions:The findings of this study suggest that the hairy roots of Ficus carica can be considered as a promising natural source of antileishmanial agents.