In our laboratory, male rats that drank 0.0014, 0.014, or 0.14 mg/ml of total pomegranate polyphenols in water for 2, 4, or 6 weeks were euthanized, perfused tran-scardially with saline, and brains were removed. The brain stems were homogenized followed by Western gel electrophoresis to measure concentrations of COX-2 (EC 18.104.22.168). Beta-actin was used to confirm that similar amounts of proteins were loaded onto each lane. Pomegranate juice consumption caused a reduction of the amount of the proinflammatory enzyme COX-2 in brain homogenates while actin content was the same in each gel (Figure 5.4). The effect was greatest after 6 weeks of diet. This suggests that pomegranate polyphenols may have anti-inflammatory effects in the brains of rats. In another study,62 anthocyanin mixtures from other plant species (Amelanchier alnifolia, A. arborea, and A. canadensis) inhibited COX-1 and -2 enzymes by 51 to 76%. The positive controls (aspirin, Celebrex, and Vioxx) inhibited COX-2 by 69 to 85%, indicating a comparable effect of anthocy-anins.62 In another study, the anthocyanins delphinidin and cyanidin inhibited LPS (lipopolysaccharide)-induced COX-2 expression in isolated macrophages, but pel-argonidin, peonidin, and malvidin did not,63 indicating a selective response. This effect was dose-dependent and likely due to blockade of MAPK-mediated pathways by anthocyanins.63 Strawberry extracts, rich in ellagic acid much like pomegranates, have been shown to inhibit COX enzymes in vitro,64 which would modulate the inflammatory process. These studies are supportive of our results and suggest that the reductions of COX-2 levels seen with pomegranates are due to the presence of anthocyanins and ellagic acid.
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