Study On The Improvement Of Malvidin-3-Galactoside On Intestinal Microbiota And The Metabolic Function In HepG2-Induced Damage Mice

The berries contain a large amount of anthocyanins with biological activity.At present,anthocyanins have been shown to have the potential to improve the disease state of the host and regulate the intestinal microecology.The study of the regulation of Malvidin-3-galactoside(M3G)on intestinal microbiota and metabolic function is unclear.With the development of high-throughput sequencing technology,more and more studies rely on high-throughput sequencing platforms to explore the connection between biologically active substances and host microorganisms.In recent years,the damage of the host body has caused the intestinal flora imbalance and other phenomena have been confirmed one after another.Therefore,to explore the regulatory effect of M3 G on the intestinal flora and its metabolic function of mice damaged by Hep G2 cells in order to find an improvement Intestinal health flora regulators or dietary supplements provide a theoretical basis.This study has explored the regulatory effects of M3 G on liver function in mice,the effect of M3 G on the composition and abundance of intestinal flora in mice,and the specificity of M3 G on the metabolic function of intestinal flora and microbial metabolism in mice.The specific conclusions are as follows:1.Measure the average feed consumption,body weight,tumor mass,tumor volume growth trend,tumor suppression rate and other basic indicators of mice.It was found that 80 mg/kg M3 G increased the average daily feed consumption of mice,significantly reduced the weight gain,the growth trend of tumor quality and tumor volume.Calculating the inhibitory rates of M3 G on tumor growth found that the inhibitory rates of 40 mg/kg M3 G,80 mg/kg M3 G and 5-fluorouracil(5-F)on tumor growth were 37.32%,46.13%,and 47.54%,respectively.The serum and liver tissues of mice were collected.The results showed that the serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)and alkaline phosphatase(ALP)levels of mice in the model group were higher than those in the model group.In addition,we also measured nitric oxide(NO),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),malondialdehyde(MDA)and inducible nitric oxide synthase(INOS)contents,the results prove that M3 G has a protective effect on liver function in Hep G2-induced damage mice.2.The 16 S r RNA high-throughput sequencing was used to analyze the regulatory effects of M3 G on the composition and abundance of intestinal microbiota in mice.Through the screening of key species,we found that 80 mg/kg M3 G had more advantages in the regulation of microbiota abundance than 40mg/kg M3 G.For example,the former promoted the enrichment of beneficial bacteria such as S24-7,Ocillospira,Ruminococcaceae,Ruminococcus,Akkermansia,Clostridiales and Prevotella in mice intestine(p < 0.05),and significantly reduced the abundance of pathogenic bacteria such as Enterobacteriaeae,Enterococcus,Actinobacteria and Clostridium(p < 0.05).At the same time,the prediction of picurst metabolic function showed that both high dose M3 G and chemotherapy drug 5-F could well regulate the abundance of microbiota closely related to the development of the disease.The results showed that the feces of model mice were enriched with a large number of opportunistic and pathogenic bacteria.In the model group,the composition and abundance of species level microbiota were positively regulated after high dose of M3 G.For example,the abundance of pathogenic bacteria Escherichia coli and Klebsieela was significantly reduced(p <0.05),while that of Clostridioides difficile was significantly enriched(p <0.05).3.The effect of M3 G on the KEGG metabolic pathway of intestinal microbiota in Hep G2-induced damage mice was analyzed by macrogenomics.The results showed that M3 G significantly improved the KEGG metabolic pathway of the tricarboxylic acid cycle by increasing the expression level of some key proteins(por A,dlat,ace E,pc and ogdh).By analyzing the Pearson correlation between protein abundance expression and microbial abundance,we found that the increased Muribaculum intestines in mice after M3 G supplementation may be one of the important factors affecting the KEGG pathway of the microbial TCA cycle.In addition,M3 G regulates the KEGG metabolic pathways of taurine,arginine and vitamin B6 by reversing the expression levels of several key functional genes(gad,roc F,sdh A/B and fadj).