Study On Liver Protective Activity Of Coriolus Versicolor In Different Developmental Stages And Its Fermentation

The fruiting body is annual and grows from summer to autumn.According to the Chinese Pharmacopoeia,the harvest period of C.versicolor is the whole year.In real life,some merchants will deliberately select the white（good looking）C.versicolor to sell at a high price,but there is no corresponding scientific basis to prove that the development stage（immature stage）C.versicolor is the best quality.In this study,the differences of chemical components in the fruiting bodies of different development stages were analyzed,and the liver-protecting activities of the fruiting bodies of different development stages and the fermentation of black beans of different development stages were studied.In the review part,the research progress of C.versicolor pharmacological action and the application of fermentation method in traditional Chinese medicine were introduced.Tissue and spore separation method was used to separate and prepare the spore strains of C.versicolor,the cultivation experiment of was carried out in the TCM laboratory.From the observation of bag cultivation,it can be seen that there are three stages:black stage in the early stage,yellow stage in the middle stage and cyan stage in the mature stage.The effect of tissue separation was better than spore separation in the cultivation process.Mixed sawdust made the primordium formation time shortest,and the side X-shaped opening was more suitable for the bag cultivation than the top one.The content of polysaccharide in cyan stage was significantly higher than that in yellow stage and extremely significantly higher than that in black stage.The content of total amino acids was still the highest in the cyan period,and the content in the yellow period was higher than that in the black period.Analysis of metabolites in the black,yellow and cyan fruiting bodies revealed the presence of 62 metabolites,14kinds of fatty acyl compounds,11 kinds of carboxylic acids and their derivatives,4kinds of organic oxygen compounds,and other compounds.There are mainly 23metabolic pathways,of which phenylalanine metabolism,nicotinic acid and nicotinamide metabolism have significant influence.This part of the study can provide reference for the study of its pharmacodynamic mechanism.Liver protecting pharmacodynamics of different development stage fruiting body was studied.Water extract low,medium,and high dose was used in this pharmacodynamic study.The results showed that the liver index of mice in each experimental group was significantly decreased,except in medium and high dose groups.The serum urea content of other water extract groups was significantly increased.Serum albumin contents were increased except for the high dose group.Serum total protein contents of mice in water extract groups increased.The effect of yellow and cyan groups on improving the level of IL-6 in the liver of mice was more significant than that of black C.versicolor group.Compared with alcohol model group,the contents of TNF-αin liver of mice were significantly decreased by water extract of C.versicolor at all three stages of development,but there was no significant difference between groups.Hepatic histopathology showed that the water extract could repair liver injury in mice.The results showed that the fruit-body of three developmental stages could repair the liver injury caused by alcohol in mice,and the effect of cyan C.versicolor medium dose group was optimum.Furthermore,results showed that there was no significant difference between the liver protection effect of the water extract of fermented black soybean and that of the water extract of fermented black soybean on alcoholism mice.The results of intestinal flora showed that,compared with the blank group,the diversity of microbial flora in feces of mice in the other experimental groups decreased,except that the community diversity of intestinal contents of mice in the water extract group was increased.The reason may be that different extracts of fermented tempeh can promote the growth of specific intestinal flora.The proportion of Lactobacillus probiotics in the intestinal flora of mice decreased from 15.63%to 5.72%after alcohol administration,while the proportion of Lactobacillus probiotics in the intestinal flora of mice was increased to28.48%after the water extract of C.versicolor,which was better than that of the positive drug group（22.03%）.It was better than by 10.23%in the water extract group of tempeh.The results indicated that the water extract of C.versicolor was better than the water extract of fermented black bean in improving the abundance of such probiotics.The results showed that the bidirectional fermentation could retain the original hepatoprotective effect.However,in the adjustment of intestinal probiotics,the water extract of C.versicolor is still more effective.The antioxidant activities of different polar extracts of fermented tempeh in vitro and their protective effects on oxidative damage of AML-12（mouse normal liver cells）induced by hydrogen peroxide were also investigated.Methane dichloromethane,ethyl acetate and propanol were extracted from the methanol extract by organic solvent extraction method.The antioxidant activities of different polarity extracts of tempeh in vitro were investigated by DPPH,ABTS and FRAP tests.AML-12 cells were induced by H₂O₂ to establish the oxidative injury model of normal mouse liver cells and to investigate the protective effects of extracts of different layers on damaged cells.The results showed that the extracts of ethyl acetate,dichloromethane and n-butanol from C.versicolor fermentation could promote the proliferation of AML-12 cells at the concentration of 5μg/m L-25μg/m L.The activities of antioxidant enzymes SOD and CAT and the contents of GSH in cell culture medium were increased in each dose group（P<0.05）,and the content of MDA in cells was decreased in each dose group.The n-butanol layer had the best protective effect on the damage of AML-12 cells induced by hydrogen peroxide,followed by the ethyl acetate layer and dichloromethane layer.When the concentration of n-butanol extract of tempeh was 25μg/m L,the apoptosis rate of cells induced by hydrogen peroxide decreased from 20.81%to 7.08%.This part of the study provides a reference for the study of dietary therapy and health care of these fermented products.