P. abrotanoides dried root has been successfully used for the treatment of Cutaneous leishmaniasis in Iranian traditional medicine as a poultice. To provide a scientific reason for ethnomedicinal use of P. abrotanoides, in this study the leishmanicidal effect of Perovskia abrotanoides extracts was evaluated on promastigotes of L. major in vitro. In this study, the antileishmanicidal effect of different extract of Perovskia abrotanoides root was evaluated on the promastigotes of Leishmania major in vitro. The dried and ground root of the plant was extracted using either maceration in 80% ethanol or Soxhlet in methanol. Then, 5 different concentrations (0.06, 0.12, 0.25, 0.5 and 1 mg/ml) of each extract, one positive control (Amphotericin B, 0.5 mg/ml), one negative control (culture medium), and one solvent control (DMSO) were prepared and were placed in a 24-well plates containing 50,000 parasites/well. The plates were incubated at 25o C for six days and the number of parasites in each well was determined on days 2, 4, and 6 of experiment microscopically using Neubauer chamber. It was observed that amphotericin B and both macerated and Soxhlet extracts at concentration of 1 mg/ml killed all the parasites. Lower doses exhibited a dose-dependent antileishmanial activity. The average of IC50 for macerated extract in DMSO was 4.03 × 10-2 mg/ml and for Soxhlet extract in DMSO was 7.33 × 10-2 mg/ml. The control solvents had no significant effect on the L. major promastigotes. These results indicated that both macerated and Soxhlet extracts of Perovskia abrotanoides have favorable leishmanicidal activity.