Peroxisome Proliferator-Activated Receptor-γ-Mediated Polarization of Macrophages in Leishmania Infection
Curcumin induces PPARγ mRNA expression and reduces iNOS mRNA expression in infected macrophages. In (a), PECs of BALB/c mice were infected with L. donovani promastigotes, and then 10 μM of curcumin (brown bar) or vehicle control (0.1% acetone) was added. After 2 days, the cells were harvested for RNA isolation to determine the level of PPARγ and β-actin expression by real-time RT-PCR. In (b), murine RAW264.7 cells were infected with L. donovani for 16–20 hours. Then, different concentrations of curcumin were added. Thirty minutes after curcumin treatment, IFNγ was added to activate the macrophages. At 5 hours after the addition of IFNγ, the cells were harvested, mRNA was extracted, and conventional RT-PCR was performed. The gel shows the end point PCR-amplified iNOS (496 bp) and β-actin cDNAs products. In (c), nonelicited PECs from resistant C3H and susceptible BALB/c strains were cultured with L. donovani promastigotes in wells that contained cover slips for a period of 16–20 hours; then curcumin and IFNγ and TNFα were added. On day 4-5, the coverslips were stained to enumerate the percent of infected macrophages and number of amastigotes per macrophage under a microscope, similar to steps described in Figure 3.
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