| Summary: | 碩士 === 國立中興大學 === 食品暨應用生物科技學系所 === 107 === Black garlic is produced resulting from chemical and biochemical reactions in the fresh garlic bulb (Allium sativum L) in a high temperature and controlled humidity condition for a few days. In particular, black garlic has strong anti-oxidant, anti-liver cancer and hypolipidemic effects as compared with fresh raw garlic. Previous studies in our laboratory had found that hot water extracts of black garlic had anti-inflammatory potential. However its anti-inflammatory ingredients and mechanisms during aging process still remain unclear. Therefore, the purpose of this study was to investigate changes in the composition of hot water extracts and anti-inflammatory effects of polysaccharides and polyphenol-rich extracts from black garlic during aging process LPS-stimulated primary peritoneal macrophages from female BALB/c mice. The results showed that polysaccharides in the black garlic aging for 20 days, (20dBGPS) inhibited the secretion of TNF-α dose-dependently. Polysaccharide in 10, 17 and 20 days significantly decreased the secretion ratio of TNF-α/TL-10 at a concentration of 40 and 200 ug/ml. The polyphenol-rich extracts with different aging days also inhibited the secretion of TNF-α. Overall of the results that revealed the black garlic aging for 20 days had the best anti-inflammatory effects. Therefore black garlic aging for 20 days was selected to investigate anti-inflammatory ingredients and mechanisms produsted of 20dBGPS and 20dBGEE were purified using gel filtration. Black garlic polysaccharides, 20dBGPS, were resolved into three factions including BPSF1, BPSF2 and BPSF3 using Sepharose 6B gel filtration column. The mean molecular weight of BPSF1 was 1.40 x 106 Da, and its monosaccharide composition was mainly galactose, xylose and mannose. The mean molecular weight of BPSF2 was 2.39 x 106 Da, and its monosaccharides were mainly composed of galactose, galacturonic acid and arabinose. The mean molecular weight of BPSF3 was 4.27 × 103 Da, and its monosaccharides mainly consisted of glucose, galactose and mannose. All three fractions had anti-inflammatory effects for they significantly reduced TNF-α secretion by LPS-stimulated macrophages. BPSF3 had the lowest mean molecular weight which the best anti-inflammatory effect. To unravel possible and anti-inflammatory mechanisms, real time quantitative polymerase chain reaction was used to evaluate changes in relative expression amounts of inflammation-related molecules. We found that BPSF3 down-regulated TLR-2 and NF-κB gene expression, suggesting that BPSF3 achieved its anti-inflammatory effects by down-regulating of NF-κB gene expression in TLR signaling pathways. However 20dBGSP and BPSF1, 2 did not share the anti-inflammation pathway. The purification of 20dBGEE using Sephadex LH-20 gel filtration resolved into three factions, BGEE1, BGEE2 and BGEE3, but they did not have anti-inflammatory effects, respectively. It is speculated that the anti-inflammatory effect of polyphenol in black garlic might through be synergistic or additive manners.
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