Effects Of Lentinan Polysaccharide On Immunomodulator Activity And Signaling Pathways PI3K/Akt And NF-κB In Mice

Lentinus edotus originated in China and is the world’s second largest mushroom.It is known as the"King of mountain delicates"and has a delicious and smooth taste.It is mostly used in the treatment of anorexia,exhaustion and deficiency of qi in Chinese traditional medicine.Lentinan is a kind of macromolecular carbohydrate with biological activity in Lentinus edodes.It has biological functions such as immune regulation,anti-tumor,antioxidant,anti-aging,anti-inflammatory and anti-virus.In order to investigate the immunomodulatory effect of Lentinan on splenic lymphocytes of mice in vitro,spleen of Balb/c mice was extracted aseptically to make splenic lymphocyte suspension.The immunosuppressed mouse model was established by cyclophosphamide induction in vivo,and the mice were given intragastric administration and intraperitoneal injection of cyclophosphamide.The immunomodulatory effects of Lentinan polysaccharide were studied through in vivo and in vitro experiments to provide theoretical and experimental basis for the development and application of lentinus edodes polysaccharide.The research methods and results are as follows:1.Primary mouse spleen lymphocytes were used as target cells and cultured in vitro for48 hours.Determined by MTT,Elisa,RT-q PCR and flow cytometry method detection of lentinan on spleen lymphocyte proliferation index,IL-6,IL-4,IL-12 and IFN-γcytokine secretion level and m RNA expression level,percentage of lymphocyte subsets.The results showed that in the dose range of 10～80μg/m L,lentinan could increase the proliferation index of mouse splenic lymphocytes,the secretion and m RNA expression levels of IL-6,IL-4,IL-12 and IFN-γ,the percentages of CD4~+and CD8~+subsets of splenic lymphocytes,and the ratio of CD4~+/CD8~+.Lentinan can increase the percentage of T cell subsets by increasing the number of single positive T cell subsets,and play a bidirectional regulatory role in the body’s immunity with low concentration promoting high concentration inhibition.2.To establish an immunosuppressed mouse model induced by cyclophosphamide,and to investigate the effect of Lentinan on the immune function of spleen lymphocytes in immunosuppressed mice in vivo.Determined by MTT,Elisa,RT-q PCR and flow cytometry method detection of lentinan on cyclophosphamide immunosuppressive mice spleen lymphocyte proliferation,IL-6,IL-4,IL-12 and IFN-γcytokine secretion level and m RNA expression level,percentage of lymphocyte subsets.The results showed that in the dose range of 10～80 mg/kg,lentina edodes could improve the immune organ index,splenic lymphocyte proliferation index,cytokine secretion and m RNA expression level,CD4~+and CD8~+percentages of splenic lymphocyte subsets and CD4~+/CD8~+ratio of cyclophosphamide induced immunosuppression mice.Lentinus edodes polysaccharide can reduce the damage of immune organs in immunosuppressed mice induced by cyclophosphamide,thereby improving the immune response of the body.3.Immunosuppressed mice were induced by cyclophosphamide western blot was used to detect the effect of lentinan on PI3K/Akt and NF-κB signaling pathways in spleen lymphocytes of immunosuppressed mice induced by cyclophosphamide.The results showed that in the dose range of 10～80 mg/kg,lentinan could increase the protein expression levels of p-PI3K,p-Akt,IKK-α,IKK-βand NF-κB in spleen lymphocytes of immunosuppressed mice.Lentinus edochis polysaccharide can play an immunomodulation role by activating PI3K/Akt and NF-κB signaling pathways.