摘要
为了研究茶薪菇(Agrocybe chaxingu Huang)液体培养胞外代谢产物体内外抗氧化特性,本研究以油茶枝与麦麸为主要原料,通过三个试验:比较不同营养基质的茶薪菇液体培养胞外代谢产物中多糖与酚类物质含量及其体外抗氧化特性,并研究对液体培养不同胞外代谢产物的分离制备技术(试验一);利用β-环状糊精包被的液体培养上清液,分析茶薪菇胞外代谢产物混合物对小鼠脏器指数与血清中抗氧化指标的影响(试验二);在此基础上,进一步分析茶薪菇胞外多糖与酚类代谢产物对小鼠生长性能、脏器指数以及血清中抗氧化指标的影响(试验三)。
分别以麦麸、油茶枝水提液培养基作为对照(CK-WB和CK-YC),与麦麸和油茶枝的混合水提液培养基(WB-YC)做比较;测定液体培养茶薪菇上清液中多糖与酚类物含量及其体外抗氧化特性;冷冻干燥醇析分离的胞外多糖与酚类产物,并分析制备产物的多糖与多酚物质含量。对茶薪菇的液体培养结果显示:上清液中,胞外多糖含量CK-WB组与WB-YC组高于CK-YC组(P<0.05),而酚类代谢物含量WB-YC组与CK-WB组高于CK-YC组(P<0.05);三组茶薪菇液体培养上清液对羟自由基的体外清除率顺序为:CK-WB > WB-YC > CK-YC(P<0.05)。
利用β-环状糊精(β-CD)包被,喷雾干燥制备茶薪菇胞外代谢产物混合粉;以4%油茶枝水提液组(CK-YC)为对照,对小鼠进行不同灌胃水平的比较试验。结果显示:灌胃不同剂量的4%油茶枝水提液(CK-YC)以及3%麦麸水提液+4%油茶枝水提液(WB-YC)的茶薪菇液体培养上清液制备胞外代谢产物,小鼠胸腺指数都较生理盐水灌胃组呈提高趋势;以灌胃600-800mg/kg·bw·d剂量的WB-YC组喷雾干燥制备产物,小鼠血清中SOD指标显著高于对照组(P<0.05)。
通过醇析分离,冷冻干燥制备茶薪菇液体培养胞外多糖与胞外酚类物质;分析其对小鼠灌胃试验的在体抗氧化特点。试验结果显示:与对照组相比,WB-YC组与CK-YC组的茶薪菇胞外酚类代谢物,能显著提高小鼠血清中的SOD活力(P<0.05),而其他各组的茶薪菇液体培养胞外多糖也不同程度地显示对提高小鼠血液中SOD活力的促进作用。同时,小鼠血液中MDA含量分析也显示,与对照组相比,不仅3%麦麸+4%油茶枝多糖组(P<0.05),其他各茶薪菇胞外代谢产物灌胃组均不同程度地降低血液中MDA含量。此外,分别灌胃茶薪菇液体培养胞外多糖与胞外酚类物,对小鼠脏器指数没有明显影响作用。
In this study, the water extracts of Camellia oleifera Abel branch powder and wheat bran were used in liquid medium for the submerged culture of Agrocybe chaxingu Huang. Three experiments were conducted, in which to compare the extracellular metabolites of A. chaxingu in polysaccharides and phenolic compounds from different culture medium and to study the techniques in the preparation for extracellular metabolites (Experiment 1); to evaluate the in vivo antioxidant effect of spray dried extracellular metabolites withβ-cyclodextrin (Experiment 2), freee dried extracellular polysaccharides and phenolic metabolites (Experiment 3), respectively, in mice. The growing performance, of mice and their viscera index were compared, the SOD and MDA content in serum were measured.
The water extract of wheat bran (CK-WB), Camellia oleifera Abel branches (CK-YC) and the mixture of the water extract of wheat bran and Camellia oleifera Abel branches (WB-YC) were compared as three culture media. The extracellular polysaccharide and phenolic compounds in the supernatant of submerged culture was separated with ethanol precipitation, then freeze dried. The content of polysaccharides and phenolic, and their in vitro antioxidant characters were determined. The results showed that the total polysaccharides in supernatant of subermerged culture was higher in the group of CK-WB and WB-YC than in CK-YC (P <0.05), while the total phenolic content was higher in WB-YC group followed by CK-WB than in CK-YC group (P <0.05). The clerarance rate of hydroxyl radical in Fention system rated in the order: CK-WB> WB-YC> CK-YC (P <0.05).
The extracellular metabolites of A. chaxingu were spray dried by use ofβ-cyclodextrin and were given to mice orally by gavage in different dosages. The results showed that the thymus index of mice was increased by giving spray dried submerged culture supernatant in all dosage level for CK-YC and WB-YC. In addition, for mice given 600-800mg/kgbw.d of spray dried supernatant of submerged culture, the SOD of mice serum was significantly higher in WB-YC than in CK-YC group (P <0.05).
Further more, the effect of phenolic and polysaccharide metabolits on improving antioxidation capacity of mice in vivo was evaluated. Comparing with mice given with saline, the extracellular phenolic metabolites of A. chaxingu in WB-YC and CK-YC groups increased SOD activity in mice serum significantly (P <0.05). The increases in SOD activity were also observed in mice give by polysaccharide metabolits. Moreover, comparing with mice given with saline, the decrease of MDA content in mice serum was found from various groups, especially in polysaccharide group of WB-YC (P<0.05).
引文
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