The aim of this study is to investigate the antioxidant and intracellular anti-inflammatory efficacy of blueberry leaf extracted with hot water (BLW), 70% ethanol (BLE), and 70% acetone (BLA) in RAW 264.7 macrophages. In order to evaluate the anti-inflammatory effect of blueberry leaf extracts, RAW 264.7 macrophages were stimulated with lipopolysaccharide (LPS) to induce the production of inflammation-related factors, which were measure by Western blotting and real-time PCR methods. i-NOS, COX-2 protein, and mRNA expression showed concentration-dependent decrease. The decreases in the mRNA expression levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE2) were concentration-dependent. Further, the antioxidant effects of blueberry leaf on total polyphenol contents, electron donating ability and ABTS+ radical scavenging activity were evaluated. The total polyphenol contents of BLW, BLE, and BLA were 217.04±2.98, 156.72±3.90, and 182.88±3.02 mg TAE/g, respectively, while the electron donating abilities at 1,000 μg/mL of BLW, BLE, and BLA were 81.7, 79.6, and 79.3%, respectively. The ABTS+ radical scavenging activity was fond to be concentration dependent. The nitric oxide (NO) production inhibition activities at 50 μg/mL of BLW, BLE, and BLA were 35.1, 42.4 and 42.7%, respectively. In conclusion, the antioxidant and anti-inflammatory test results indicate that blueberry leaf extracts (BLW, BLE, and BLA) can be used as potential anti-inflammatory agents.

The nutritional composition and in vitro anti-oxidant activities of blueberry (Vaccinium ashei) leaf extract were investigated to examine their physiological characteristics. Calcium was the most abundant mineral. The principal free sugars were glucose, sucrose, maltose, and fructose. The amino acids were mainly composed of glutamic acid and aspartic acid. The fatty acids consisted mainly of 40.94% saturated fatty acid and 54.35% unsaturated fatty acid. In addition, the 112.64 mg% of vitamin C was analyzed as a natural anti-oxidant. Based on the bioactivity-guided isolation principle, the resulting ethanolic extracts from the blueberry leaf were divided into several fractions of n-hexane, chloroform, ethyl acetate, and water. The ethyl acetate fraction showed the greatest total phenolic content. The total phenolics and flavonoid were 50.51 mg of GAE /g and 13.09 mg%, respectively. The ABTS-radical-scavenging activity of the ethyl acetate fraction was 97.53% at a concentration of 500 μg/mL. The ferric-reducing anti-oxidant power of the ethyl acetate fraction increased in a dose-dependent manner. The results suggest that the ethyl acetate fraction of the blueberry leaf extract has good in vitro anti-oxidant activities and excellent nourishment, and can thus be useful food resources.