Analysis Of Chemical Composition Of Leucocalocybe Mongolica And Study On Its Pharmacological Activities And Mechanism

In this paper,the tumor inhibition experiments were carried out on different components(LMP,TPLM,ELM,SLM,and two sterol compounds)of L.mongolica,and their mechanism of action was discussed,as well as the study of high-throughput blood non-targeted metabolomics and the regulation of intestinal flora.On the other hand,the total polysaccharides,sterols and water extracts(LMP,SLM,WLM)of L.mongolica were researched on disorder of blood glucose metabolism.The nutrient components of L.mongolica were analyzed by kjeldahl method,amino acid automatic analysis method,soxhlet extraction method,inductively coupled plasma atomic emission spectrometry and burning method.The results showed that the total protein content of L.mongolica was 43.56% in total protein,35.58% in total polysaccharide,9.2% in total ash and 1.09% in crude fiber.The proportion of essential amino acids required by human body in L.mongolica was significantly higher than that of several common edible fungi in the market.The heavy metal content of L.mongolica is lower,and it is a first-grade edible product which was safety for hunman being.The GC-MS and LS-MS methods were used to analyze the chemical composition of the fruiting body of L.mongolica.The results of GC-MS analysis showed that a total of 31 chemical components with a matching degree higher than 85% were detected,and all the detected compounds were mainly volatile aliphatic compounds with less polarity.Through the result of LS-MS analysis,a total of 50 compounds were obtained,including 9 phenolic compounds,12 fatty acid compounds,3 nucleotide compounds,8 quinone compounds,6 carbohydrate components and steroids There are 4 kinds of compounds in several components,and there are 8 other kinds of compounds.Isolation and extraction of sterols from Mongolian Pleurotus ostreatus using forward silica gel and Sephadex LH-20 column chromatography,and Ergosterol peroxides,Ergosterol,(22E,24R)-Ergostere-7,22-diene-3,5,6,Ergoster-triol,ergoster-4,6,8(14),22(23)-tetramene-3-ketone and other 5 sterol compounds were isolated.This study has a certain understanding of the composition of total sterol of L.mongolica.Due to the unreasonable anthropogenic collection of L.mongolica and the deterioration of environmental conditions,the habitat of L.mongolica has been destroyed and the species is on the verge of extinction.Therefore,the method of deep fermentation was used to produce its fermentation mycelium in large quantities,which protected the resources of L.mongolica to some extent.On the basis of previous researches,the conditions of fermentation and culture in high-density fermenting tanks were optimized,and the final results were obtained: the medium formula: soybean powder 16.55g/L,corn pulp powder 33g/L,sucrose was 33g/L under the initial conditions,and gradually increased to 9% with the fermentation time.Fermentation conditions: under the optimized conditions of 10 L fermentation tank,the initial p H was adjusted to 6.5 under the condition of 25?,the mechanical rotor speed of the fermentation tank was 150 RPM,and the initial inoculation amount was 12.5 %,which was the best culture conditions for the fermentation tank.The amplification culture was conducted in 50 L and 100 L fermentation tanks under these conditions.Final results: at 216 h,the biomass of 50 L fermenters reached the maximum,17.84g/L.The biomass of 100 L fermentation tank reached 19.64g/L at 192 h.With the amplification of fermentation tank,the fermentation period can be obviously reduced,which provides a basis for industrial production of liquid fermentation of L.mongolica.The chemical components of the fermentation mycelium were compared with those of the fruiting body,and a total of 31 compounds were obtained from the analysis of the fermentation mycelium,including phenols,sugars,quinones,nucleotides,fatty acids,sterols and other components.The experimental results confirmed that the mycelium of L.mongolica fermentation could replace the fruiting body for production and application to some extent.Most of the compounds are shared with fruiting bodies.Anti-tumor experiments of different components of L.mongolica were carried out by visceral index and tumor inhibition rate calculation,ELISA,H&E,TUNEL,immunohistochemical fluorescence analysis and western blotting.Total protein(TPLM),total polysaccharides(LMP),small subclass compounds extracted from formic acid(ELM),total sterols(SLM)compounds and ergostere-4,6,8(14),22(23)-tetraene-3-ketone,(22E,24R)-ergostere-7,22-diene-3,5,and 6,were used for antitumor experiments in vivo.All the experimental results showed that the above components of L.mongolica had obvious inhibition on tumor inhibition of H22 tumour-bearing mice,among which,SLM and total LMP of L.mongolica were the best.High-throughput nontargeted metabolomics and regulation of intestinal flora were studied in SLM.The experimental results show that after SLM treatment,the tumor microenvironment,compared with model group,affect the body's metabolic disorder produced strong disturbance of compounds in tryptophan metabolic pathways,serotonin can synaptic way,way of protein digestion and absorption,linoleic acid metabolic pathways,the biosynthetic pathways of amino acid and arachidonic acid metabolism pathway in several metabolic pathways,such as obvious callback state,has the tumor inhibition performance.And intestinal microbial changes.After SLM treatment,Prevotella and related genus has been proved to promote lipid metabolism,treat metabolic syndrome,cancer and other diseases.In addition,Phascolarctobacterium,Bacteroidaceae and Bacteroides are also enriched in SLM group,which play a variety of important roles in maintaining human health and provide essential nutrition for the body.Research on the regulation of blood sugar disorders.According to the literature,L.mongolica is used in the treatment of T2DM.The author of this paper applied hypoglycemic experiments on fasting blood glucose,ELISA,glucose tolerance test and Western Blot method for the total polysaccharide(LMP),water extract(WLM)and total sterol group(SLM)of L.mongolica.The experimental results showed that both LMP and WLM had better hypoglycemic effects,and 4 weeks of treatment could significantly improve glucose and lipid metabolism and insulin resistance.While the hypoglycemic effect of SLM group was not obvious.These results suggest that LMP and WLM can be used as active components in the prevention and treatment of T2DM.