Effect Of Fermented Barley Extract On Lipometabolism In 3T3-L1 Preadipocytes And Its Mechanism

People with obesity resulting to abnormal lipid metabolism are quick exceeding in recent years, so that world health organization(WHO) has defined obesity as an world-wide infection disease. Therefore, screening natural biochemical with anti-obesity function from natural food has become a hotspot on preventing and curing obesity. Barley is a common and wide-spread cereal in our country and is called as complete-value food for concluding wide nutritional and bioactive components. It has been proven that barley contains many antioxidant, anti-aging,anti-hyperlipidenmia and anti-diabetes bioactive ingredients, including beta-glucose,flavone, polyphenol and so on. Microorganic biotransformation effectively improves the content of anti-obesity components in fermented barley, so that the way not only provides a new and creative experimental theory for developing reducing-lipid natural product, but also provides an effective way to use barley.In my paper, raw barley extract(RBE), Lactobacillus plantarum dy-1 fermented barley extract(LFBE) and Saccharomycetes fermented barley extract(SFBE) were prepared with barley as fermentation substrate. Researched the bioactive components with reducing-lipid function in fermented barley extract by measuring the main component content of unfermented and fermented barley extract; Analyzed the effect of fermented barley extract on inhibiting the viability and differentiation of preadipocytes in vitro and discussed the reducing-lipid mechanism. The following was the main results:1. The content of main components of unfermented and fermented barley extract were compared and analyzed. The content of protein, polyphenol, polysaccharide andβ-glucan of unfermented and fermented barley extract were determined by national stander ways; SDS-PAGE analysis technology was used to analyze the change of protein molecular mass between unfermented and fermented barley extract; High performance liquid chromatography(HPLC) analysis technology was used to analyze the content of varied phenolic acid in unfermented and fermented barley extract. The results showed that polysaccharide contents in LFBE and SFBE were reducedcompared to RBE; The content of β-glucose in LFBE was improved from 4.83% to12.80%, but β-glucose in SFBE was reduced to 2.01%. Protein and polyphenol contents were significantly improved after fermentation both in LFBE and SFBE;Protein in 30~40 kDa was significantly increased in LFBE compared to SFBE and RBE. Both lactobacillus plantarum dy-1 and saccharomycetes fermentation had a significant effect on the content of polyphenol, including gallic acid, cumaric acid,protocatechuic acid and vanillic acid; Ferulic acid in LFBE was significantly increased, but was reduced in SFBE.2. The effect of fermented barley extract with different cultures on 3T3-L1 preadipocytes viability was researched. The effect of fermented barley extract on viability of preadipocytes was evaluated by CCK-8 method. Compared to RBE, both LFBE and SFBE significantly inhibited the viability of 3T3-L1 preadipocytes;Inhibitory rates of LFBE and SFBE in 500 μg/mL on intervening 3T3-L1 preadipocytes for 24 h were 61.69% and 21.94%, and the inhibitory rates were 66.96%and 20.80% for 48h; LFBE had a better inhibitory effect than SFBE, and its half-inhibitory concentration(IC50) was 560μg/mL.3. The effect of fermented barley extract with different cultures and its main bioactive ingredients on differentiation of 3T3-L1 preadipocytes was researched. The effect of fermented barley extract on differentiation of preadipocytes was evaluated by oil red O staining. The results shown that RBE ineffectively inhibited the differentiation of 3T3-L1 preadipocytes, but LFBE had an effectively inhibitory effect which had a positive correlation with the content; LFBE had a better inhibitory rate than SFBE because that their inhibitory rates in 400 μg/mL were 58.85% and 12.53%,respectively. Protein and polysaccharide in LFBE had an effectively inhibitory effect on differentiation of 3T3-L1 preadipocytes, and their inhibitory rates were 34.95%and 10.49%, respectively; However, Protein and polysaccharide in SFEB ineffectively inhibited the differentiation of 3T3-L1 preadipocytes. Polyphenol in LFBE and SFBE were the main bioactive ingredients with inhibitory effect on differentiation of 3T3-L1 preadipocytes, and their inhibitory rates were 15.38 % and 9.16%, respectively;Ferulic acid and vanillic acid were the main ingredients in polyphenol with inhibitoryeffect on the differentiation of 3T3-L1 preadipocytes.4. The mechanism under the inhibitory effect of LFBE on differentiation of3T3-L1 preadipocytes was researched. The expression levels of crucial genes about lipid mechanism were analyzed by Real-time PCR method. The results showed that LFBE and its protein, polyphenol compounds and ferulic acid all significantly inhibited the expression of crucial genes about lipid mechanism, including C/EBPα,PPAR-γ, SREBP-1c, PTP1 B and aP2. The comprehensive effect resulted in inhibiting lipid accumulation and reducing of the amount and size of lipid dropt in 3T3-L1 preadipocytes.