Apoptotic Effect of Morphine, Imiquimod and Nalmefene on Promastigote, Infected and Uninfected Macrophages with Amastigote of Leishmania Major by Flow Cytometry

The parasites of genus Leishmania are the causative agents of one of the most widespread and devastating diseases. According to follow-up data, these medications may provoke adverse drug reactions, drug resistance, relapse, as well as financial burden. The mechanism of action of opioid drugs is primarily exerted via transmembrane G-protein coupled receptors. One of the potent synthetic immunomodulator agents is imiquimod with low molecular weight and unknown mechanism of action. Monocyte and macrophage are the primary site of action for imiquimod. Nalmefene is a well-known opioid antagonist agent which simultaneously inhibits these receptors and augments intracellular pathogenicity, hence providing opportunities to investigate their function. The aim of present work was evaluating the effect of morphine, imiquimod and nalmephen on the Leishmania major and investigating cytotoxic effect this drug on the uninfected macrophage and infected macrophage for detected early apoptosis, necrosis and secondry apoptosis by flowcytometry method. In this study we used morphine, imiquimod, nalmefene, and Glucantime. We treated promastigotes, macrophages, and infected macrophages with above drugs, and the apoptosis evaluated by flow cytometry. The results showed that in all concentration of morphine more than 98% of promastigotes remained alive that it is deduced that morphine lacks any lethal effect on L. major after 24 h, whereas in groups treated with Glucantime alone or in combination with Nalmephene and Imiquimod, 84.13%, 88.96% and 86.72% of promastigotes were alive, respectively. The results of macrophage treatment with morphine, imiquimod, and nalmefene demonstrated that most necrosis has occurred in nalmefene group (6.54%).