| First of all, the species of wild mushroom in Taihang Mountains of Hebei Province were investigated and their ecological characteristic was analyzed in this thesis. Secondly, the extracts from four kinds of wild mushrooms, including Suillus luteus, Coprinus micaceus, Coriolus hirsutus and Trametes pubescens, were prepared by using different pola organic solvent. Also, four kinds of polysaccharide were obtained via using hot water extraction and ethanol precipitation. The antioxidant active component of each organic extracts was analyzed and the antioxidative activity of every extracts was measured in vitro or vivo.The results were as follows:1. During last three years, 145 species wild mushrooms were investigated in Taihang Mountains in rainy season (from July to October every year). These fungi belonged to 2 subphylums, which are Basidiomycotina and Ascomycotina. The Basidiomycotina included 137 species from 70 genera, 26 families, 7 orders, and 3 classes. The Ascomycotina included 8 species from 7 genera, 5 families, 3 orders and 2 classes. Among the mushrooms, 55 species belonged to edible fungus, 38 species belonged to medicinal fungus, 13 species belonged to toxic fungus, 6 species belonged to ectomycorrhizal fungus and 47 species belonged to saprophytic fungus.2. The extracts from Suillus luteus, Coprinus micaceus, Coriolus hirsutus and Trametes pubescens, were obtained by using different pola organic solvent, including petroleum ether (PE), ethylacetate (EA) and ethanol (Eth). Then polysaccharides from four kinds of mushrooms were obtained by using hot water extraction and ethanol precipitation.(1) The contents of antioxidant component, including polyphenols, triterpenes and flavonoids were measured. As a result, there were different polyphenols and triterpenes among every organic extracts from four fruitbodies, including PE-soluble portion, EA-soluble portion and Eth-soluble portion. The content of polyphenols involved in PE-soluble portion, EA-soluble portion and Eth-soluble portion of Suillus luteus, which is 4.62%, 6.4% and 4.8%, respectively, is more than that in other organic extracts.The contents of triterpenes existed in PE-soluble portion and EA-soluble portion are more than that in Eth-soluble portion from four fruitbodies. The content of triterpenes existed in the PE-soluble portion from four fruitbodies varied from 20.35% to 23.68%. The content of triterpenes in EA-soluble portion from Suillus luteus or Coriolus hirsutus is 19.18% and 24.8%, respectively, which are the most in four fruitbodies. The most content of triterpenes in Eth-soluble portion from Coriolus hirsutus is 14.4%, and Suillus luteus is 8.59% in the next. The most content of flavonoids of three organic extracts of Suillus luteus is 1.7%, 4.92% and 0.43%, respectively. However, the PE-soluble portion, Eth-soluble portion from Coprinus micaceus and PE-soluble portion from Coriolus hirsutus have no flavonoids.(2) The antioxidant assays of every extracts in vitro were carried out by four methods, including DPPH radical-scavenging method, inhibition of lipid peroxidation, determination of reducing power and metal ion chelating activity. The results showed that each extracts from four kinds of fruitbodies exhibited antioxidant activity in vitro.The IC50 value of DPPH radical-scavenging assays of the three kinds of organic extracts of Suillus luteus was 0.00015μg/mL, 0.18 mg/mL and 0.26 mg/mL, respectively, and lower than that of other mushroom organic extracts and polysaccharide portion. The IC50 value of EA-soluble portion and Eth-soluble portion from Coriolus hirsutus and Trametes pubescens varied from 1.27 mg/mL to 1.60 mg/mL, also had better ability in DPPH radical-scavenging. The PE-soluble portion of Coprinus micaceus had the highest IC50 value of 23.08 mg/mL.The organic extracts from four fruitbodies were more effective than their polysaccharides in inhibition of lipid peroxidation. The IC50 value of three organic extracts of Suillus luteus was 0.04 mg/mL, 0.08 mg/mL, 0.14 mg/mL, respectively. The value three organic extracts from Trametes pubescens was 0.14 mg/mL, 0.13 mg/mL, 0.31 mg/mL, respectively. The value of the PE-soluble portion and EA-soluble portion from Coprinus micaceus was 0.13 mg/mL and 0.31 mg/mL in turn.In metal chelating activity assay, the Eth-soluble portion and polysaccharides from each fruitbodies had better chelating activity than other extracts. The Eth-soluble portion and polysaccharide of Suillus luteus, and polysaccharide of Coprinus micaceus had the lowest IC50 values, which were 2.78 mg/mL, 1.44 mg/mL and 0.88 mg/mL, respectively. The EA-soluble portion of Suillus luteus had the highest reducing power with a OD values of 0.479 at 10 mg/mL. Also, the PE-soluble portion, Eth-soluble portion from Suillus luteus and EA-soluble portion of Coriolus hirsutus exhibited higher reducing power than other extracts, the OD value was 0.342, 0.26, 0.325, respectively.According to the integrated analysis of IC50 value obtained by above methods, the Eth-soluble portion of Suillus luteus, Coprinus micaceus and Trametes pubescens, EA-soluble portion and Eth-soluble portion of Coriolus hirsutus all exhibited better antioxidant activity.Furthermore, the antioxidant activity of organic extracts from Suillus luteus was better than that from other fruitbodies.3. Aged mice caused by D-galactose were perfused by Eth-soluble portion of Suillus luteus using ascorbic acid (Vc) as control. The results showed that the Eth-soluble portion of Suillus luteus could increase the activities of SOD and GSH-px and decrease the content of MDA in serum and liver of mice. The effect of the extracts was equal to or better than that of ascorbic acid. Under the treatment of large dose of Eth-soluble portion of Suillus luteus, the activity of SOD would be increased to 467.7 U/mL in serum and 41.02 U/mL in liver of mice. Also, the activity of GSH-px would be increased to 385.56 unit in serum and 3.15 unit in liver. Compared to normal group and aged group, the activities of SOD and GSH-px were significantly increased. The content of MDA in serum and liver were significantly decreased to 3.65 nmol/mL and 0.33 nmol/mgprot, respectively. These suggested that the Eth-soluble portion of Suillus luteus could increase the antioxidant ability of aged mice in vivo. |
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