| Camellia seeds belong to the mature seeds of Camellia Oleifera Abel.Camellia seed contain a variety of bioactive substances,including unsaturated fatty acids,flavonoids,polyphenols,saponins,polysaccharides and proteins,which have bioactive functions such as antioxidant,anti-inflammatory,hypoglycaemic and hypolipidaemic.The study of anti-inflammatory activity is the most prominent.In this paper,UPLC-MS and microbiomics techniques were used to investigate the anti-enteritis activity of ethanolic extract of Camellia seeds and its mechanism of action,and to explore monomer compounds with anti-enteritis activity,providing a theoretical basis for the development of enhanced efficiency of Camellia Oleifera Abel oil.(1)The bioactivity of ethanol extract from 15 Camellia seed samples was determined.The results showed that the inhibition rate of Camellia seed ethanol extract on LOX activity was 74.28%-96.74%.The inhibition rate of COX-2 activity was between 6.06%and 84.85%.The inhibition rate of α-glucosidase activity was 47.33%-99.64%.The inhibition rate of α-amylase activity was 2.8%-59.36%.The inhibition rate of pancrelipase activity was between 51.76%and 95.06%.The lipid oxidation inhibition rate ranged from 66.13%-88.12%,hydroxyl radical scavenging rate ranged from 9.75%-71.98%,and superoxide anion scavenging rate ranged from 8.93%-58.74%.The results of Drosophila Smurf experiment showed that ethanolic extract of Camellia seed inhibited enteritis in the range of 15.06%-61.71%,while the drug control of salazosulfapyridine was 27.99%.The differences between varieties were significant.The correlation results suggest that inhibition of LOX activity may be one of the anti-enteritis mechanisms of the ethanolic extract of Camellia seeds.(2)The LC-MS technique was used to detect small molecule metabolites in 15 ethanolic extracts of Camellia seeds and 5 plant polyphenol extracts(Cinnamon polyphenol,Apple polyphenol,Grape skin polyphenol,Pomegranate peel polyphenol,Echinacea polyphenol).885 metabolites were identified,carbohydrate metabolites had the highest relative content,accounting for 30.25%of the total metabolites;followed by polyphenols,lipids and amino acid compounds.Correlation analysis,molecular docking and Drosophila Smurf assay revealed that N,N-dimethylarginine dihydrochloride,Naringenin chalcone,Isomangiferin,Wighteone and Kaempferitrin exhibited high anti-enteritis activity.It was verified that Kaempferitrin and Wighteone have good inhibitory effect on LOX,COX-2 and Drosophila enteritis.The IC50 for Kaempferitrin inhibition of LOX,COX-2 and Drosophila enteritis were 0.1295 mmol/L,0.1860 mmol/L and 0.3651 mmol/L,respectively;The IC50 for LOX,COX-2 and Drosophila enteritis inhibition by Wighteone was 0.2156 mmol/L,0.2789 mmol/L and 0.4243 mmol/L,respectively.(3)The effects of different Camellia seeds ethanol extracts on gut microbiota of DSS-induced Drosophila enteritis were studied by 16S rRNA sequencing.The results of microbial colony composition,alpha-diversity and beta-diversity showed that the effect of ethanolic extract of Camellia seed on the intestinal microflora of Drosophila was mainly manifested by altering the abundance of intestinal flora and increasing the ratio of Firmicutes/Bacteroidetes.Analysis of the correlation between gut microbiota with anti-inflammatory activity and the relative content of Camellia Oleifera Abel seed metabolites revealed that modulation of the abundance of Enterococcus,Dysgonomonas and Sphingobacterium may be one of the mechanisms underlying the anti-enteritis activity of ethanolic extracts of Camellia Oleifera Abel seeds.Six,eight and three metabolites were significantly negatively correlated with Enterococcus,Dysgonomonas and Sphingobacterium respectively,limonin,protocatechuic,enol-phenylpyruvat and coniferylaldehyde were the highest metabolites. |
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