Gene Expression Profiles of Inducible Nitric Oxide Synthase and Cytokines in Leishmania major-Infected Macrophage-Like RAW 264.7 Cells Treated with Gallic Acid

Abstract

The effects of gallic acid on the gene expressions of inducible nitric oxide synthase (iNOS) and the cytokines interleukin (IL)-1, IL-10, IL-12, IL-18, TNF-α, and interferon (IFN)-γ were investigated by reverse-transcription polymerase chain reaction (RT-PCR). The experiments were performed in parallel in non-infected and in L. major-infected RAW 264.7 cells and the expression profiles were compared with those mediated by IFN-γ plus lipopolysaccharide (LPS). The infection per se induced the expression first of IL-1 and TNF-α mRNA, later that of IL-10 mRNA. Gallic acid induced low and transient levels of TNF-α and IL-10 in non-infected cells, and it clearly enhanced and prolonged iNOS and cytokine mRNA expressions in Leishmania-parasitised cells. Interestingly, and in contrast to activation by IFN-γ/LPS, gallic acid also stimulated Leishmania-infected cells to produce IFN-γ mRNA. For IFN-α, a sandwich immunoassay was performed to determine its amount present in the supernatant of gallic acid-stimulated RAW 264.7 cells. In showing predominant stimulation of infected cells and the induction especially of IFN-γ, a cytokine that plays a central role in antimicrobial macrophage and T cell regulation, these data provide the basis for an immunological concept of gallic acid and possibly other plant polyphenols for their beneficial effects in various infectious conditions.

Key words

Leishmania major - Macrophages - gallic acid - inducible nitric oxide synthase - cytokines

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