Isolation And Extraction Of Polysaccharides From Five Edible Fungi And Their Effects On Immune Function In Mouse

Polysaccharide is one of the main active ingredients of edible fungi.It has rich effect and is widely used in various biomedical and functional foods.Edible fungi polysaccharides have a variety of biological activities,such as immune activity,antitumor and antioxidant activities and hypoglycemic activities,among which the most prominent is the immune activity.At present,there are a lot of edible fungi polysaccharides with immune activity in the stage of research and development,but whether there are differences in the immune effects of different edible fungi polysaccharides is not completely clear.Therefore,five edible fungi were selected as research objects in this study,and their water-soluble polysaccharides were isolated and extracted.In this study,five edible fungi that have been widely reported to have immune-enhancing effects were selected(Inonotus obliquus、Phellinus igniarius、Lentinula Edodes、Hericium erinaceus and Agaricus blazei Murill)as a research object,the water-soluble polysaccharides were isolated and extracted,and on this basis,immunosuppressive mouse models were established to study the immune activities of five edible fungal polysaccharides,and to preliminarily reveal the immune activities of different edible fungal polysaccharides and their possible mechanisms.Specific research contents are as follows:(1)Water-soluble crude polysaccharides were extracted from Fruiting bodies of five edible fungi,including Inonotus obliquus,、Phellinus igniarius,、Lentinula Edodes,、Hericium erinaceus and Agaricus blazei Murill,.Polysaccharide content and protein content were determined by phenol sulfuric acid method and Coomassie brilliant blue method.The results showed that the extraction rates of water-soluble polysaccharides from five edible fungi were respectively Inonotus obliquus 1.058 %、Phellinus igniarius 2.122 %、Lentinula Edodes 4.048 %,、Hericium erinaceus 6.556 % and Agaricus blazei Murill 5.034 %.Sevage method was used to remove proteins from crude polysaccharides,After removing protein,the content of Inonotus obliquus polysaccharide was 83.55%、the polysaccharide content of Phellinus igniarius is 85.51 %、Lentinula Edodes contains 69.71 % polysaccharide、the polysaccharide content of Hericium erinaceus was 56.98 % and the polysaccharide content of Agaricus blazei Murill was72.51 %..After purification,the protein content of polysaccharides: Inonotus obliquus polysaccharides 3.45 % 、 Phellinus igniarius polysaccharides 1.4 % 、 Lentinula Edodes polysaccharides 1.34 %、Hericium erinaceus polysaccharides 0.72 % and Agaricus blazei Murill polysaccharides,1.21 %.(2)Cyclophosphamide intraperitoneal injection was used to establish an immunosuppressive mouse model,and the isolated and extracted Inonotus obliquus polysaccharide(200 mg/kg),Phellinus igniarius polysaccharide(200 mg/kg),Lentinula Edodes polysaccharide(200 mg/kg),Hericium erinaceus polysaccharide(200 mg/kg),and Agaricus blazei Murill polysaccharide(200 mg/kg)were used to intervene mouse for 12 days,and on the8 th,9th and 10 th days,a mouse immunosuppression model was established with a dose of 40mg/kg cyclophosphamide,and the body weight,weight of mouse in different treatment groups were observed.Index and tissue morphological changes of the spleen and thymus;Real-time PCR was used to verify the key genes of DNA repair and cell DNA damage response biological processes.Flow cytometry was used to detect the analysis of mouse thymus and splenic T lymphocyte subsets and apoptosis,and the immune activity of different polysaccharides was evaluated.The results showed that Inonotus obliquus polysaccharide,Phellinus igniarius polysaccharide,Lentinula Edodes polysaccharide,Hericium erinaceus polysaccharide,and Agaricus blazei Murill polysaccharide After intervention,spleen coefficients of mouse were increased by 0.6±0.02 g,、0.36±0.18 g,、0.68±0.02 g,、0.46±0.26 g and 0.54±0.3g.Compared with the model group,the spleen coefficients of mouse after the intervention of birch Inonotus obliquu polysaccharide,、Phellinus igniarius polysaccharide,、Lentinula Edodes polysaccharide、Hericium erinaceus polysaccharide and Agaricus blazei Murill polysaccharide,increased by1.08±0.11 g,、0.62±0.02 g,、0.57±0.19 g,、0.33±0.06 g and 0.22±0g,respectively.。The spleen structure and cell morphology of mouse were observed by HE staining.The results showed that the spleen cell morphology and cell density were more complete and higher after the intervention of Inonotus obliquus polysaccharide、Phellinus igniarius polysaccharide、Lentinula Edodes polysaccharide,and Agaricus blazei Murill polysaccharide,and the boundary between red pulp and white pulp was clear.These results indicated that Inonotus obliquus polysaccharides,、 Phellinus igniarius polysaccharide,、 Lentinula Edodes polysaccharide,、Hericium erinaceus polysaccharide and Agaricus blazei Murill.polysaccharides could effectively improve the spleen injury caused by cyclophosphamide in mouse and enhance the immune function of the body.The results of T-lymphocyte subsets and apoptosis in the thymus and spleen of mouse showed that;Inonotus obliquus polysaccharide 、 Phellinus igniarius polysaccharide、Lentinula Edodes polysaccharide and Agaricus blazei Murill polysaccharide both of them could effectively protect T cells from the effects of cyclophosphamide;Inonotus obliquus polysaccharide、Phellinus igniarius polysaccharide、Lentinula Edodes polysaccharide,and Agaricus blazei Murill polysaccharide all of them could effectively protect mouse spleen T cells from the effects of cyclophosphamide.The proportion of apoptosis in spleen cells of Inonotus obliquus polysaccharide group、Phellinus igniarius polysaccharide group、Lentinula Edodes polysaccharide group、Hericium erinaceus polysaccharide group and Agaricus blazei Murill polysaccharide group decreased significantly,and the anti-apoptosis effect of Inonotus obliquus polysaccharide group and Hericium erinaceus polysaccharide group was more significant..The apoptotic ratio of thymus cells in Inonotus obliquus polysaccharide、Phellinus igniarius polysaccharide、Lentinula Edodes polysaccharide,Hericium erinaceus polysaccharide and Agaricus blazei Murilli polysaccharide groups decreased.The results of real-time real-time PCR detection showed that Lentinula Edodes polysaccharide Edodes could significantly reduce the gene expression of Mrel1 and 53BP1,and that Inonotus obliquus polysaccharide,、Phellinus igniarius polysaccharide,、 Hericium erinaceus polysaccharide,and Agaricus blazei Murill polysaccharide could significantly increase the expression of Nek1 gene.This study will provide experimental basis for functional research and product development of edible fungal polysaccharides.