To investigate neuronal cell protective effects of an ethyl acetate fraction from chestnut inner skin, in vitro assays, including 2 0 ,7 0 -dichlorofluorescein diacetate, 3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT), and lactate dehydrogenase (LDH), were performed. Intracellular accumulation of reactive oxygen species resulting from hydrogen peroxide (H2O2) treatment of PC12 cells was significantly reduced when ethyl acetate fractions were present in the medium compared to PC12 cells treated with H2O2 only. In a cell viability assay using MTT, the ethyl acetate fraction protected against H2O2-induced neurotoxicity, and inhibited LDH release into the medium. In addition, the ethyl acetate fraction improved in vivo cognitive ability against amyloid b-peptide (Ab)-induced neuronal deficit. High-performance liquid chromatography analyses showed that gallic acid, catechin, and epicatechin were predominant phenolics in the ethyl acetate fraction. Consequently, the results suggest that chestnut inner skin, including above phenolics, could ameliorate Ab-induced learning and memory deficiency, and be utilized as effective substances for neurodegenerative disorders, notably Alzheimer's disease.

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.

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