| As a traditional ethnic medicine,there has been a lot of researches on Tricholoma mongolicum,but there is no research on its immune function of small molecular compounds of T.mongolicum so far.In order to comprehensively study its medicinal value,based on consulting a large number of literature,this paper studies the pharmacological effects of different extracts of T.mongolicum’s fruit body in the treatment of mice with low immunity,and further explores the mechanism of T.mongolicum Objective to investigate the therapeutic effect of the small molecular part of T.giganteum on immunocompromised mice,and to analyze,isolate and identify the chemical components of the small molecular part.The research contents are as follows:The Summary part is as follows: the pharmacological activities and chemical constituents of T.mongolicum at home and abroad were summarized.The chemical constituents of T.mongolicum were mainly limited to polysaccharides and sterols.Modern pharmacological studies have found that T.mongolicum fruiting body,extract and crude polysaccharide have anti-tumor,immune enhancement,anti-oxidation,blood pressure and hypoglycemic effects.T.mongolicum has been recorded in ancient and modern materia medica.It is cold in nature and sweet in taste.Meridian tropism of the T.mongolicum was predominated by the tropism of spleen meridian or stomach meridian.It has the effect of promoting intestines and benefiting spleen and stomach.Therefore,the chemical composition and immune activity of T.mongolicum were studied on the basis of review.The research part is as follows:1.Resource investigation and collection of T.mongolicum: through three years of field investigation,visiting local herdsmen and market investigation.It is known that the harvest season of T.mongolicum is mostly in summer and autumn.Through local herdsmen,it is known that the edible and medicinal methods of T.mongolicum,and because it can improve the body’s immunity,its price is also increasing year by year.Microscopic identification and molecular identification were used to identify the authenticity of the collected T.mongolicum.2.Study on the immune activity of crude extract in vivo:Prevention and treatment of low immunity of mice through administrating fruiting body extract of T.Mongolia were investigated.The Immunosuppressive Model of mice was established by intraperitoneal injection of cyclophosphamide.To explore the therapeutic effects of the high,medium and low dose groups of T.Mongolia fruiting body petroleum ether extract and ethyl acetate extract on immunodeficiency.The weight of mice was measured,interleukin 2(IL-2),interleukin 4(IL-4),interferon-γ(IFN-γ),immunoglobulin A(Ig A)indexes were detected,and the spleen was analyzed,Pathological changes of the thymus.The results showed that high dose of petroleum ether extract(30mg / kg)could significantly improve the spleen index of mice,improve the atrophy of thymus and spleen caused by cyclophosphamide,enhance the secretion of cytokines and the phagocytic capacity of cells,maintain the normal serum biochemical indexes,and protect the immune organs The effect of the dose group is close to that of levamisole hydrochloride,which is a positive drug.It has the potential of development and utilization,and the mechanism needs to be further studied.3.Studies on chemical constituents:In order to determine the active ingredient of the treatment of hypoimmunity in small molecule fraction,the chemical constituents of small molecule fraction were studied.High-performance liquid chromatography,preparative liquid chromatography and Nuclear magnetic resonance were used to analyze,separate and identify the chemical constituents of small molecule fraction.Finally,seven compounds were obtained,which were ergosta-7,22-dien-3βylpalmitate(1),4,6,8(14),22,(23)-tetraen-3-one-ergostane(2),β-Sitosterol(3),Ergosterol(4),Ergosterol peroxide(5),Ergosta-7,22-dien-3β—ol(6),Cerevisterol(7).Through gc-ms analysis of petroleum ether small polar components,the majority of fatty acids in petroleum ether extract of T.mongolicum,and the content of oleic acid was the highest.Among them,the isolated compounds 4,6,8(14),22,(23)-tetraen-3-one ergostane,β-sitosterol,ergosterol,ergosterol peroxide have the highest yield for subsequent use experiment.4.In vitro immunoactivity of steroids: RAW264.7 cells induced by LPS were used to screen the activity of the four compounds in vitro,and in vitro experiments were conducted to further explore the therapeutic effect of small molecules on immunosuppression.The results showed that the four compounds could significantly inhibit the proliferation of RAW264.7 cells induced by LPS,and promote the release of no and the secretion of IL-1 β,IL-6 and TNF-α cytokines.The effect of 4,6,8(14),22,(23)-tetraen-3-one ergosterone was the most obvious,and the cell morphology and intercellular junctions after administration were very similar to those in the blank group.Compared with the blank control group,te at the best effective concentration promoted the phagocytosis of RAW264.7 cells by 0.51 times.5.Study on the in vivo immune activity of steroids: Study on the immunosuppressive effect of T.mongolicum steroids,and further verify the therapeutic effect of small molecule compounds on immunosuppressive mice.The results showed that steroids can significantly enhance the organ index of immunosuppressive mice,promote the secretion of cytokines IL-2,IL-4,Ig A and IFN-γ,and increase the production of hemolysin.6.In conclusion,this paper studies the positive effect of traditional Chinese medicine Mongolian Mushroom on the spleen,stomach and body through in vivo and in vitro munological activity test.And the paper determines the basis of its active substances as steroids.The above research results have been verified by in vivo and in vitro pharmacological experiments,providing a scientific basis for further researches. |
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