포도주 부산물의 화학성분 분석 및 생리활성

Abstract

This study analyzed flavonoid and phenol contents and fatty acid composition of wine by-product and investigated biological activities of crude extracts and fractions of wine by-product. The flavonoid and phenol contents of the methanol (MeOH) extract were greater than those of the acetone+methylene chloride (A+M) extract. Among fractions, the 85% aqueous methanol (85% aq. MeOH) fraction contained the highest flavonoid contents, while the n-buthanol (n-BuOH) fraction had more phenol contents. In fatty acid composition, the percentages of linoleic acid, linolenic acid and oleic acid were 47.8%, 3.3% and 5.98%, respectively.

In anticancer activity of wine by-product, treatments of crude extracts and fractions significantly inhibited the growth of HT-29, AGS and HT-1080 human cancer cell lines (p<0.05). The A+M and MeOH extracts showed a inhibitory effect on growth by more than 50% at the concentrations of 0.5 mg/mL. Among the fractions, the 85% aq. MeOH and n-Hexane fractions exhibited a higher inhibitory effect on proliferation of three types of cancer cells.

In antioxidant activity of wine by-product, all tested extracts and fractions dose-dependently decreased cellular reactive oxygen species (ROS) production induced by H202 in comparison with that produced by exposure to the extract-free control. The MeOH extract showed a higher inhibitory effect on cellular ROS producing than that of the A+M extract at all concentrations tested. Treatment with the BuOH fraction (0.1 mg/mL concentration) inhibited cellular ROS production by 60% (p<0.05). In DPPH and ABTS radical scavenging assay, the MeOH extract showed a scavenging effect greater than that of the A+M extract (p<0.05). The n-BuOH fraction showed stronger radical inhibitory effect than dibutyl hydroxy toluene (BHT). The A+M and MeOH extracts from wine by-product significantly inhibited genomic DNA oxidation (p<0.05). The 85% aq. MeOH and n-BuOH fractions showed a higher DNA oxidation effect compared with control (p<0.05). In addition, the crude extracts and fractions increased levels of intracellular glutathione (GSH) in a dose manner.

In anti-inflammatory activity, the productions of nitric oxide (NO) assay showed that the extracts and fractions significantly reduced NO production induced by lipopolysaccharide (LPS) (p<0.05). The n-BuOH fraction showed the highest reduction of NO production by 61.3% and then A+M and MeOH extracts showed reduced NO production by 52.3% and 50.1%, respectively (p<0.05). In cytokine production assay, the treatment with 85% aq. MeOH fraction (3 ㎍/mL concentration) significantly reduced concanavalin A (Con A)-induced IL-2 expression at 6 h and LPS-induced IL-2 expression at 48 h, as compared to treatment with stimulant (Con A or LPS) (p<0.05). IL-6 expression after 48 h of treatment with LPS was significantly reduced by 85% aq. MeOH fraction (10 ㎍/mL) (p<0.05). The 85% aq. MeOH fraction (10 ㎍/mL) with LPS reduced IL-12/IL-20(p40) expression after 24 h and 72 h of treatment (p<0.05). The 85% aq. MeOH fraction plus Con A significantly increased IL-4 (3 ㎍/mL) and IFN-γ(10 ㎍/mL) expression at 24 h (p<0.05).

These results suggested that 85% aq. MeOH and n-BuOH fractions of wine by-product inhibited cellular oxidation and growth of human cancer cells and their activity may be associated with the contents of flavonoids and phenols. Thus, the recycling of winery co-products or side streams constitutes an opportunity for providing valuable materials to the pharmaceutical, cosmetic nutraceutical, and food industries, contributing to reductions in both the costs and environmental impact linked to the disposal of these by-products in the production areas.