The pine bark extract was added during the beer manufacturing process to develop a functional beer using natural plant materials. Pilsner beer was prepared by adding 0-0.125% to the wort. Immediately before fermentation, extracts were added with yeast cultured twice for 48 hr, and fermentation was carried out at 25°C for 5 d and storage was carried out at 2°C for 15 d. In the experiment, pH, reducing sugars, soluble solid contents, specific gravity, alcohol contents, chromaticity, total flavonoids, total polyphenols, DPPH, and yeast number were tested during fermentation and storage to characterize. The pH, reducing sugar, soluble solid contents, specific gravity, and alcohol contents of the six samples were not significantly different during fermentation and storage. However, yeast number, flavonoids, polyphenols, and DPPH were higher at higher pine bark extract contents. L value in chromaticity was the highest in control, and a value and b value were higher as the amount of extract added. In the sensory test, beer with 0.05% pine bark extract showed the best color, sweetness and aroma. Therefore, it was judged that it is desirable to prepare beer by adding 0.05% pine bark extract to pilsner beer.

Amyloid-β protein (Aβ) is known to increase free radical production in neuronal cells, leading to cell death by oxidative stress. The purpose of this study was to evaluate the protective effects of PineXol® on Aβ25-35 induced neuronal cell death. Rat pheochromocytoma (PC-12) cells were pre-treated with 100 μg/mL of PineXol® for 2 h. The cells were exposed to single dose of 30 μM Aβ25-35 for 24 h. Cell death was assessed by a cell count kit-8 (CCK-8) assay, lactate and dehydrogenase (LDH) release assay. An Apoptotic process was analyzed by a protein expression of the Bcl-2 family using western blotting. Cell viability increased in PC-12 cells treated with both Aβ25-35 and PineXol®, compared to the control group. PineXol® induced a decrease of the Bcl-2 protein expression (p<0.05), while Bax and Sod1 increased (p<0.05), indicating attenuation of Aβ25-35 induced apoptosis. These results suggest that PineXol® may be a good candidate for the prevention of Alzheimer’s disease(AD).

The purpose of this study was to evaluate the protective effect of PineXol® on H2O2-induced cell death in SK-N-MC cells, and in early stage focal ischemia rodent model. SK-N-MC cells were pre-treated with 200 μM H2O2 or various concentrations of PineXol® (10, 30, and 50 pg/mL) for 24 h, and then exposed to H2O2 for 3 h. Cell death was assessed by the CCK-8 assay, reactive oxygen species (ROS) assay, and lactate and dehydrogenase (LDH) release assay. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) expressions were also analyzed by western blotting. Focal ischemia rodent model was used as the in vivo model, and different concentrations of PineXol® (1, 10, and 100 mg/kg) were administered. One week after administration, reduction of infarct volume was analyzed by TTC staining. Cell viability of H2O2-treated SK-N-MC cells significantly increased by pre-treatment of PineXol® (p<0.05). PineXol® pre-treatment also induced significant decrease of ROS and LDH expressions. However, PineXol® did not affect the infarct volume. These results suggest that PineXol® has significant neuroprotective effect in vitro, but statistical significance was not confirmed in the in vivo focal ischemia mo