In vitro and in vivo Assessment of Anti-Leishmanial Efficacy of Leaf, Fruit, and Fractions of Juniperus excelsa Against Axenic Amastigotes of Leishmania major and Topical Formulation in BALB/c Mice

Background: Leishmaniasis is a parasitic disease caused by Leishmania protozoa. Iran is an endemic region for leishmaniasis and thus, many natural medicaments, such as Juniperus excelsa, are traditionally being used for the treatment of its cutaneous form. Objectives: The current study aimed at assessing the anti-leishmanial activities of this medicament’s leaf and fruit extracts and respective leaf fractions against Leishmania major as the causative agent of zoonotic cutaneous leishmaniasis in both in vitro and in vivo models. Methods: This experimental study was carried out at Shiraz University of Medical Sciences, Shiraz, Iran, during the year 2013. For the generation of axenic amastigotes, promastigotes were mass cultivated and incubated at 33 to 34°C and pH 3.5. The anti-amastigote activity was evaluated using the colorimetric assay. For in vivo study, promastigotes were inoculated in 40 female BALB/c mice tails. Lesions were created after four weeks. Animals were randomly distributed to four groups as test, placebo, Glucantime, and control. Mice were treated for 30 days. Lesion diameters were measured and recorded weekly. Results: All extracts and fractions possessed potent activities; leaves showed the highest with IC50 (SD) (50% inhibitory concentration) 0.97 3.53 mg/mL, and among fractions, ethyl acetate with IC50 1.95 5.30 had greater anti-leishmanial activity. In animal models compared with the beginning, the diameter of Cutaneous Leishmaniasis (CL) lesion had a significant difference in the test group (P < 0.05 vs. control group) at the end of the protocol. However, no significant differences were seen in placebo and Glucantime groups versus the control group. Conclusions: Comprehensive and long term therapy assessment with J. excelsa is needed to introduce it as a natural antileishmanial medicine.