IN VITRO EFFECTS OF WARBURGIA UGANDENSIS, PSIADIA PUNCTULATA AND CHASMANTHERA DEPENDENS ON LEISHMANIA MAJOR PROMASTIGOTES

Abstract

Plant extracts from Warburgia ugandensis Sprague (Family: Canellaceae), Psiadia punctulata Vatke (Family: Compositae) and Chasmanthera dependens Hoschst (Family: Menispermaceae) were tested for activity on Leishmania major promastigotes (Strain IDU/KE/83 = NLB-144) and infected macrophages in vitro. Plants were collected from Baringo district, dried, extracted, weighed and tested for antileishmanial activity. Serial dilutions of the crude extracts were assayed for their activity against Leishmania major in cell free cultures and in infected macrophages in vitro. Inhibitory concentrations and levels of cytotoxicity were determined. Warburgia ugandensis, Psiadia punctulata and Chasmanthera dependens had an IC50 of 1.114 mg/ml, 2.216 mg/ml and 4.648 mg/ml, respectively. The cytotoxicity of the drugs on BALB/c peritoneal macrophage cells was insignificant as compared to the highly toxic drug of choice Pentostam®. The supernatants from control and Leishmania infected macrophages were analyzed for their nitrite contents by Griess reaction and nitrite absorbance measured at 540 nm. Warburgia ugandensis (stem bark water extract), Chasmanthera dependens (stem bark water extract) and Psiadia punctulata (stem bark methanol extract) produced 112.3%, 94% and 88.5% more nitric oxide than the untreated infected macrophages respectively. Plant crude extracts had significant (p<0.05) anti-leishmanial and immunomodulative effects but insignificant cytotoxic effects at 1mg/ml concentration. All experiments were performed in triplicate. Statistical analysis of the differences between mean values obtained from the experimental group compared to the controls was done by students’t test. ANOVA was used to determine the differences between the various treatment groups. The analysis program Probit was used to determine IC50s.