| The author adopts the use of research methods of literature search and review of papers regarding macro fungi in Inner Mongolia over the past twenty years for this dissertation. Accordingly, papers regarding990species of macro fungi were collected. The diversity catalogue of these990species was arranged according to Ainsworth&Bisby's Dictionary of the Fungi (2008,10th), and they belong to2phylums,23orders,78families,218genus.The survey of the flora diversity study indicated that macro fungi species are vey plentiful in Inner Mongolia. Thirteen different families each have over20species represented, comprising a total of686species, or69.29%of the total macro fungi in Inner Mongolia. Another24families have over10species represented, totaling to484species, or48.88%of the total. Of these species,531have either edible or medicinal characteristics, or both. Three hundred sixteen species have medicinal properties,373species are edible, and158species are both edible and have medicinal functions.As a result of conducting field sampling in Hexigten Banner, a total of41specimens were collected. Twenty-six species were identified through studying their gross features and their structures under a microscope. Among these26species,2phylums were identified. The first phylum was Ascomycota, containing2orders,2families,2genuses and2species. The second phylum identified was Basidiomycota, which contained7orders,14 families,20genuses and24species.Because of the diversity of the species and their unique chemical structures, the higher fungi are an extremely important resource for new natural medicines. In order to fully exploit the fungal resources in Inner Mongolia region and search for new active compounds, we investigated the chemical constituents and bioactivities of the five higher fungi species of Lenzites betulina, Ramaria flava, Agaricus spissicaulis, Geastrum fimbriatum and Leucopaxillus giganteus in Inner Mongolia region.Forty-nine different compounds, including1new compound and1new natural compound, have been isolated by the use of chromatographic methods. The structures of these compounds were elucidated by both spectroscopic methods (MS,1D&2D-NMR, IR, UV) and chemical methods, and the results are as follows:(1) Fourteen compounds were isolated from the extract of Lenzites betulina including seven sterides, three p-terphenyls, one pyranone, one amino acid, one aromatic compound and one D-allitol.(2) Eleven compounds were isolated from the extract of Ramaria flava including six sterides, one glucose, one base, two aromatic compounds and one D-allitol. All of these were first reported from the genus Ramaria.(3) This was the first time to investigate the chemical constituents of the Agaricus spissicaulis fruiting bodies. Ten compounds were isolated from its extract including four sterides, one cerebroside, one base, two coumarin compounds, one aliphatic acid and one D-allitol. (4) It was the first time to investigate the chemical constituents of the fruiting body of Geastrum fimbriatum. Subsequently, six compounds were isolated from its extract including two sterides, two glucose, one amino acid and one D-allitol.(5) Eight compounds were isolated from the extract of Lenzites betulina including two sterides, two acidic compounds, one hydroquinol, one alkaloid and one picolinic acid. All of these compounds, except the alkaloid, were first reported from this fungus.The antioxidant activities of the ethanol extracts of these five fungi and three p-terphenyls compounds were evaluated by DPPH radical scavenging activity and hydroxyl radical scavenging ability. Also, the antibacterial and antifungal activities of the extracts from these species were evaluated. Moreover, we also evaluated the in vitro cytotoxicity of these extracts against human tumor cells MDA-MB-231,BGC-803和NCI-H520, by the use of MTT assay. The results are as follows:(1) There were significant differences among the extracts of these five fungi (P<0.05) on DPPH and hydroxyl radical scavenging ability. The radical scavenging activity of these five fungi decreased in the order of Lenzites betulina, Ramaria flava, Leucopaxillus giganteus, Geastrum fimbriatum and Agaricus spissicaulis. Lenzites betulina displayed the best activity among these fungi in regard to DPPH and hydroxyl radical scavenging ability with the lowest IC50values (24.12±0.61μg/mL and24.58±0.19μg/mL respectively). The content of total phenol for these five fungi, in decreasing order, were17.79±0.57μg/mg (Lenzites betulina),12.95±0.41μg/mg (Ramaria, flava),11.66±0.56μg/mg (Leucopaxillus giganteus),10.53±0.53Hg/mg (Geastrum fimbriatum), and3.32±0.45μg/mg (Agaricus spissicaulis). The results indicated that the total phenols were positively correlated with DPPH and hydroxyl radical scavenging ability, displaying correlation coefficients of0.856and0.892, respectively. The three p-terphenyls compounds were positively correlated with DPPH and hydroxyl radical scavenging ability, and the compound2displayed higher hydroxyl radical scavenging activity (IC50=37.69±0.8μg/mL).(2) Using the inhibition zone method, we isolated the ethanol extracts from Lenzites betulina, Ramaria flava, Leucopaxillus giganteus and Agaricus spissicaulis. Lenzites betulina has an inhibitor effect on Staphylococcus aureus, Escherichia coli and Bacillus subtilis. Among these four fungi, Leucopaxillus giganteus displayed the best antibacterial activity. The inhibition zone diameters of this species were all greater than2cm, and the minimum inhibitory concentration of this species were all6.25mg/mL. The antibacterial activities of Lenzites betulina were better, and the inhibition zone diameters of this species were2.24cm,1.90cm and1.72cm respectively, with the MIC of6.25mg/mL,12.50mg/mL and25.00mg/mL respectively.Using the fungus cake method, antifungal activities of the extracts from these five species were evaluated. First, we found that the inhibition ratios of these five species on Gibberella zeae are higher than the inhibition ratios on Fusarium graminearum and Cercosporella albo-maculans. Second, it was discovered that Leucopaxillus giganteus has the best antifungi activities among the five fungi. At the concentration of1mg/mL, the inhibition ratios of Leucopaxillus giganteus on Gibberella zeae, Fusarium graminearum and Cercosporella albo-maculans were35.80%,67.23%,52.05%respectively.(3) The MTT test revealed that the ethanol extracts from these five mushrooms exhibited certain cytotoxicities against the human cancer cells identified as MDA-MB-231, BGC-803and NCI-H520. The IC50values of these five ethanol extracts also have positive correlation within this experiment scope of concentration. Of these five species, Leucopaxillus giganteus displayed the best cytotoxicities. At the concentration of200ug/mL, the inhibition ratios of Leucopaxillus giganteus against MDA-MB-231, BGC-803and NCI-H520cells were77.35%,68.92%,65.53%respectively, with corresponding IC50values of46.26μg/mL,65.06μg/mL and84.76μg/mL. At the concentration of200μg/mL, the inhibition ratios of Lenzites betulina against MDA-MB-231,BGC-803and NCI-H520cells were83.15%,55.54%and57.50%respectively, with corresponding IC50values of51.47μg/mL,130.19μg/mL and106.54μg/mL.
|
PDF Full Text Request