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BackgroundIn vitro antioxidant activities and neuron-like PC12 cell protective effects of solvent fractions from aged garlic extracts were investigated to evaluate their anti-amnesic functions. Ethyl acetate fractions of aged garlic had higher total phenolics than other fractions.MethodsAntioxidant activities of ethyl acetate fractions from aged garlic were examined using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) and malondialdehyde (MDA) inhibitory effect using mouse whole brain homogenates. Levels of cellular oxidative stress as reactive oxygen species (ROS) accumulation were measured using 2',7'-dichlorofluorescein diacetate (DCF-DA). PC12 cell viability was investigated by 3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) and lactate dehydtrogenase (LDH) assay. The learning and memory impairment in institute of cancer research (ICR) mice was induced by neurotoxic amyloid beta protein (Aβ) to investigate in vivo anti-amnesic effects of aged garlic extracts by using Y-maze and passive avoidance tests.ResultsWe discovered that ethyl acetate fractions showed the highest ABTS radical scavenging activity and MDA inhibitory effect. Intracellular ROS accumulation resulting from Aβ treatment in PC12 cells was significantly reduced when ethyl acetate fractions were presented in the medium compare to PC12 cells which was only treated with Aβ only. Ethyl acetate fractions from aged garlic extracts showed protection against Aβ-induced neurotoxicity. Pre-administration with aged garlic extracts attenuated Aβ-induced learning and memory deficits in both in vivo tests.ConclusionsOur findings suggest that aged garlic extracts with antioxidant activities may improve cognitive impairment against Aβ-induced neuronal deficit, and possess a wide range of beneficial activities for neurodegenerative disorders, notably Alzheimer's disease (AD).

The ethylacetate (EtOAc) fraction of blueberry leaf extract was investigated to examine the in vivo antiamnesic effects against amyloid β protein (Aβ)-induced learning and memory deficit. The fraction showed the highest antioxidant activities, and the generation of intracellular reactive oxygen species was significantly decreased. Cell viability assays revealed the in vitro cytoprotective effects of the fraction, and the cytoplasmic lactate dehydrogenase release into the medium was dose-dependently inhibited. In addition, a chlorogenic acid was identified as a predominant phenolic compound by high-performance liquid chromatography analysis. Antiamnesic effects were evaluated by using in vivo the Y-maze and passive avoidance tests, and preadministration of the fraction attenuated Aβ-induced memory impairment in both in vivo experiments. Acetylcholinesterase prepared from mice brain was inhibited by the fraction, and malondialdehyde generation in the brain homogenate was also decreased. These findings suggest that the EtOAc fraction of blueberry leaf extract could possess a wide range of physiological effects against neurodegenerative diseases.

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.