Locked nucleic acid -anti- let-7a induces apoptosis and necrosis in macrophages infected with Leishmania major

Abstract

Background: Protozoan parasites of the genus Leishmania are etiologic agents which are intracellular pathogens of vertebrates and replicate inside infected macrophages. Leishmania have developed complex strategies to reverse host immune responses in favor of it. One of the major species causing cutaneous involvements is Leishmania major. MicroRNAs (miRNA) are non-coding small RNAs encoding 22-nucleotide (nt) long RNAs. miRNAs affect diverse biological processes, including cell cycle, proliferation, differentiation, growth and development, metabolism, aging, apoptosis, gene expression and immune regulation. This study aimed at evaluating apoptosis and necrosis after transfection locked nucleic acid (LNA) inhibitor of let-7a in the human macrophages miRNAs upon infectionwith L. major. Materials and methods: Inhibition of let-7a in macrophages was derived originally from the human monocytes (MDM), using locked nucleic acid (LNA) antagomir. The total cellular RNA was extracted 24 and 48 h post transfection. The levels o Let-7a expression was measured by qPCR Real Time using specific primer. Annexin-V/Propidium Iodide staining method was performed to detect apoptosis and necrosis in the MDM cells. Data were analyzed using the Kruskal-Wallis and Mann-Whitney tests. Results: Let-7a inhibition increased the MDM cells apoptosis and necrosis using flow cytometry method. Conclusions: The results suggested that inhibition of let-7a could be a new approach in treatment of leishmaniasis.