Comparative Study On The Chemical Composition And Anti-inflammatory Activity Of Schisandra And Its Fermented Products

Objective:Summarize the research status of Schisandra and its fermented products,determine the end of lignan synthesis of Schisandra based on transcriptome sequencing,screen fermentation strains based on total lignan,and compare the chemical composition and anti-inflammatory activity of Schisandra and its fermentation products.Provide theoretical support for the development of fermented products of Schisandra chinensis.Methods:1.we sequenced the full-length transcriptome of Schisandra chinensis in different growth periods to study the gene expression of key enzymes in the synthesis of lignans in Schisandra chinensis and the occurrence of variable splicing events;2.Based on the total lignan content,this article compares the effects of S.schisandra fermented by Saccharomyces cerevisiae,Lactobacillus plantarum and Lactobacillus paracasei on its total lignan content;3.Investigate the material-to-liquid ratio of different fermentations,time of fermentation and inoculation amount of strains,and optimize the fermentaing process of Schisandra by Saccharomyces cerevisiae;4.The content of total polysaccharides,total polyphenols and total lignans of Schisandra before and after fermentating were compared by ultraviolet spectrophotometry.The HPLC method with acetonitrile(A)-water(B)as the mobile phase was used to detect the content of seven lignan monomer components in Schisandra chinensis,and the methodology was investigated;5.Establish Quality evaluation mode to detect the content of lignan monomer,calculate the relative correction factors of the other 6 components with Schisandra A as the reference peak,and compare the detection results of the external standard method and the one-test-multiple-evaluation method;6.In order to find different compounds,the content of total polysaccharides,7.total polyphenols,total lignans and lignan monomer components in Schisandra chinensis before and after fermentation were used for principal component analysis(PCA)and least square discriminant analysis(OPLS).-DA);8.RAW264.7 macrophages containing LPS(1 μg/ml)were used to establish an inflammation model.The administration group was given 20 and 60 mg/mL of the alcohol extract of Schisandra chinensis before fermentation,and 20,60 mg/mL of the alcohol extract of Schisandra chinensis after fermentation.Detect the secretion of inflammatory mediators IL-6,IL-1β,and TNF-α.q-PCR was used to detect the expression of mRNA of IL-6,IL-1β,TNF-α,NF-κB and TLR4.Compare the anti-inflammatory effects of the alcohol extract of Beiwuweizi before and after fermentation.Results:1.Variable splicing events and the expression of key enzyme genes for lignan synthesis:CCR gene expression decreases,CAD and PLR gene expression is up-regulated from young fruit to green fruit,and then down-regulated.The number of variable shearing events in red fruit and mature fruit is greater than that of young fruit and green fruit.2.Screening strains:Saccharomyces cerevisiae has a higher total lignan content after fermenting Schisandra schisandra than before fermenting schisandra.The total lignan content of Schisandra fermented by Lactobacillus plantarum and Lactobacillus paracasei was lower than that before fermentation.3.Investigate different fermentation material-liquid ratios,fermentation time,and the amount of bacteria planted to optimize the fermentation process of S.schisandra by Saccharomyces cerevisiae,and determine the best process as follows:distillation volume 5%,material-liquid ratio 0.2 g/mL,fermentation time 15 days,the initial pH of 7.0 is the best fermentation process.4.Compare the changes in total lignans,total polysaccharides,and total polyphenols of Schisandra chinensis before and after fermentating by Saccharomyces5.cerevisiae:After fermentation,the total polysaccharide content is significantly reduced,and the reduction rate is 7.00～16.00%;the total polyphenol content is significantly increased,the improvement rate is in the range of 6.00～13.00%;the total lignan content is significantly increased,and the improvement rate is 12.00～17.00%.6.The results of one test and multiple evaluations show that the relative correction factors of the other six components calculated with Schisandrin A as the reference material are 2.771,1.108,2.390,3.186,2.688,2.286,respectively.There is no significant difference between the measured value of the external standard method and the calculated value of one measurement and multiple evaluations.7.Compare the changes in the content of 7 monomer lignans before and after fermentation of Schisandra chinensis by S.cerevisiae:The contents of schisandrin A,schisandrin A,schisandrin A,and schisandrin B increased by 15.30%,1.80%,8.02%,and 8.73%,respectively.The content of gomisin D and schisandrin B decreased by 0.95%and 1.8%respectively.The content of Schisandrin C did not change significantly,with a decrease of 0.74%.8.After modeling,the levels of inflammatory factors IL-6,IL-1β,and TNF-α in macrophage RAW264.7 increased,and the corresponding mRNA,NF-κB,and TLR-4 mRNA expression increased significantly,which was significantly different from the blank group(P<0.05).All indicators were improved after the administration intervention.At the same dosage,the schisandra extract after fermentation had better regulation of inflammatory factors than before fermentation.Conclusion:At the ripe fruit stage,the key enzyme genes are low expressed,and the synthesis of lignans is completed.At this time,it is considered as the best harvest time.After Saccharomyces cerevisiae fermented Schisandra,the content of polyphenols,total lignans,schisandrol A,schisandrin A,schisandrin B,and Schisantherin A were all in addition to polysaccharides,gomisin D,schisandrol B,and schisandrin A.raised.