Peroxisome Proliferator-Activated Receptor-γ-Mediated Polarization of Macrophages in Leishmania Infection
Activation of PPAR enhances parasitic infection. Mice were infected with L. donovani. One group was fed with 0.2 mL freshly prepared curcumin solution, given by oral gavage every other day throughout the course of the study. Curcumin was dissolved to a solution of 7.52 mg/mL in 0.1 N NaOH and immediately brought to pH 7.2 by diluting to a concentration of 11.1 μg/mL in PBS. The second group received phosphate buffered saline (PBS) in the same manner. At 4 weeks, the peak of hepatic infection (as shown in Figure 2), livers, and spleens were harvested for DNA and RNA extractions. PPAR and iNOS expression were determined by real-time RT-PCR and normalized to 18S RNA. Parasite load in the liver and spleen was determined by the real-time PCR procedure which was that of Nicolas and colleagues , except the reaction occurred in SYBR Green I PCR master mix (from Superarray). DNA, at 40 ng per reaction, was denatured at 95°C for 10 min, and then Leishmania kinetoplast DNA (kDNA) was amplified in a thermal cycler (Rotor-gene 6.0, from Corbett). The number of copies of kDNA per μg of DNA was determined using a standard curve that was established with the cloned PCR product. Filled diamonds were curcumin-treated, and clear circles were saline controls. N indicates the number of mice in each group. Statistic analysis was performed by ANOVA after the data underwent natural log transformation as described in Adapala and Chan . A value of <0.05 was considered as significant.
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