Effects Of Schistosoma Japonicum On Its Main Active Constituents By Endophytic Fungi

Schisandra chinensis(Turcz.)Baill is fruit of Magnoliaceae, which is identified as National level-3endangered medicinal plants. Fermented Chinese Herbs by microorganism can increase pharmacology of Chinese Herbs, reducing side effects and expanding indication. It is significant for saving decreasing Chinese Herbs and protecting endangered wild species.In this study, S. chinensis was chosen as substrate which was fermented by S. chinensis endophytic fungi WJl, WT4, GP5and TP2, respectively. The strain which had high ability of imporving the contents of4kinds of lignans obviously was obtained. After the selected strain was identified, the process of fermenting S. chinensis was optimized. In the research, the4kinds of lignans were also extracted and separated from non-fermented S. chinensis and fermented products. In conclusions,1.S. chinensis was fermented by single strain and mixed strains with S. chinensis endophytic fungi WJl, WT4, GP5and TP2. The results showed that the contents of4kinds of lignans in fermented S. chinensis by WJ1was obviously higher than the others, So the S. chinensis endophytic fungi WJ1was selected as research object.2. S. chinensis endophytic fungi WJ1was identified as Aspergillus niger based on its morphologic characteristics and18SrDNA molecular identification.3. The process of fermenting S. chinensis by WJl was optimized using single-factor test combined with orthogonal experimental design with the the contents of4kinds of lignans in fermented S. chinensis as markers. The optimal process conditions were as follows:the substrate concentration was0.3g/mL, the added volunm of WJ1suspension was5mL, temperature was set at30℃, the fermentation time was15d. Under the optimal conditions, the contents of schisandrin, schisantherin, deoxyschizardrin and γ-schisandrin in S. chinensis were all obviously higher than those which were obtained from crude drug, the growth rates were7.55%,3.42%,13.42%,9.86%respectively.4. The4kinds of lignans in non-fermented S. chinensis and fermented product were extracted, separated and purified. Using heat reflux with80%ethanol to extract lignans, and silica column chromatograph, macroporous resin column chromatograph to purify lignans, HPLC to analyze. In this way, schisandrin714.3684mg, schisantherin98.4508mg, deoxyschizardrin168.3487mg, and γ-schisandrin613.4260mg in fermented S. chinensis were got, and their purity were87%,90%,93%,96%, respectively. As a contrast, the4kinds of lignans from non-fermented S. chinensis were isolated with the same methods. Then schisandrin540.7900mg, schisantherin78.4923mg, deoxyschizardrin148.0257mg, γ-schisandrin471.4521mg were obtained, and their purity were87%,91%,93%,95%respectively.