Calophyllum brasiliense Modulates the Immune Response and Promotes Leishmania amazonensis Intracellular Death

Figure 4

Expression levels of cytokine and iNOS mRNA by macrophages treated with hydroalcoholic extract of C. brasiliense and infected with Leishmania (L.) amazonensis. (a) L.L.a.: L. (L.) amazonensis; C.b.: C. brasiliense. Peritoneal macrophages from BALB/c mice were treated with hydroalcoholic extract of C. brasiliense (30 μg/mL) for 3 h and infected with promastigotes of L. (L.) amazonensis (10 parasites : 1 macrophage) by more than 3 h. Macrophages were only infected with promastigotes of L. (L.) amazonensis (10 : 1) and untreated. As a negative control, macrophages were incubated only with RPMI for 6 h. significantly compared to negative control; compared to infected and untreated macrophages. (b) Cytokine mRNA expression by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) DNA fragments was separated in 1.5% agarose gel electrophoresis and revealed with ethidium bromide in a transilluminator. M: molecular marker; C−: amplification negative control (H₂O); C+: DNA obtained from macrophages stimulated with 5 μg/mL of LPS; MO: macrophages (negative control, uninfected and untreated); 4 and C.b. + L.L.a.: macrophages treated with C. brasiliense for 3 h and infected with L. (L.) amazonenis; 5 and L.L.a.: macrophages infected with L. (L.) amazonensis; 6: macrophages uninfected and untreated.

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