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Anti-inflammatory Effects of Aronia Extract on Rat Endotoxin-Induced Uveitis

¹From the Departments of Ophthalmology and Visual Sciences and ²Biochemistry, Hokkaido University Graduate School of Medicine, Sapporo, Japan; and the ³Department of Fermentative Food, Hokkaido Food Processing Research Center, Hokkaido, Japan.

PURPOSE. Aronia crude extract (ACE) with high levels of polyphenol compounds has been reported to have antioxidative effects in vitro and in vivo. In this study, attention was focused on the antioxidant effect of ACE. The purpose of the present study was to investigate the effect of ACE on endotoxin-induced uveitis (EIU) in rats. In addition, the endotoxin-induced expression of the inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 proteins was investigated in a mouse macrophage cell line (RAW 264.7) treated with ACE in vitro, to clarify the anti-inflammatory effect.

METHODS. EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). Immediately after the LPS inoculation, 1, 10, or 100 mg ACE or 10 mg prednisolone was injected intravenously. After 24 hours, the aqueous humor was collected from both eyes, and the number of infiltrating cells, protein concentration, nitric oxide (NO), prostaglandin (PG)-E2, and TNF- levels in the aqueous humor were determined. RAW 264.7 cells treated with various concentrations of ACE were incubated with 10 µg/mL LPS for 24 hours. Levels of NO, PGE2, and TNF- were determined by an enzyme-linked immunosorbent assay. The expression of iNOS and COX-2 proteins was analyzed by Western blot analysis.

RESULTS. The number of inflammatory cells, the protein concentrations, and the levels of NO, PGE2, and TNF- in the aqueous humor in the groups treated with ACE were significantly decreased in a dose-dependent manner. In addition, the anti-inflammatory effect of 100 mg ACE was as strong as that of 10 mg prednisolone. The anti-inflammatory action of ACE was stronger than that of either quercetin or anthocyanin administered alone. ACE also suppressed LPS-induced iNOS and COX-2 protein expressions in RAW 264.7 cells in vitro in a dose-dependent manner.

CONCLUSIONS. The results suggest that ACE has a dose-dependent anti–ocular inflammatory effect that is due to the direct blocking of the expression of the iNOS and COX-2 enzymes and leads to the suppression of the production of NO, PGE2, and TNF-.

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