Dracocephalum kotschyi Boiss. In Vitro Efficacy on Growth and Apoptosis Induction in Leishmania major Promastigotes.

Abstract

Dracocephalum kotschyi Boiss. is a plant generally used in modern medicine to treat many human illnesses. It is also used to prevent tumor cell proliferation throughout the world. This study's objective was to evaluate this plant's in vitro efficacy on growth and apoptosis induction in Leishmania major promastigotes. To do this, the essential oil is extracted for the test following the collection and identification of D. kotschyi. The essential oil was analyzed using a GC-MS analyzer. Promastigotes of L. major were cultured in RPMI-1640 media, and the MTT assay and a flow cytometry analysis were carried out on promastigotes that had entered the log phase. To differentiate between viable, necrotic, and apoptotic treated or untreated promastigotes, the flow cytometry method of double staining with annexin V-FLUOS and propidium iodide (PI) was used. Given the results obtained, 11 phytochemicals were identified in the essential oil of this plant. Copaene (22.15%), methyl geranate (16.31%), geranial (13.78%), and carvone (11.34%) were the main substances. The essential oil of D. kotschyi inhibits the proliferation of L. major promastigotes at 921 μg/mL, 252 μg/mL, and 416 μg/mL, respectively, after 24 h, 48 h, and 78 h. The cells were divided into four quadrates based on cell phases using the flow cytometry approach by double staining with annexin V-FLUOS and propidium iodide (PI): necrosis (Q1), late apoptosis (Q2), early apoptosis (Q3), and viable (Q4) quadrates. Overall, it is apparent that the different concentrations induced cell apoptosis in promastigotes. Observation under the light microscope at ×100 magnification showed that the different doses of D. kotschyi essential oil caused apparent alterations in the treated promastigotes. In this work, D. kotschyi essential oils induce programmed death on L. major promastigotes. This study opens many research perspectives, such as investigating the mechanisms of action and the production of a phytomedicine based on this plant.