Regulation Effect Of Antrodia Camphorata Submerged Fermented Polysaccharide On Intestinal Flora In Mice With Antibiotic-associated Diarrhea

Antrodia cinnamomea is a precious and scarce edible and medicinal fungus.This fungus belongs to Antrodia,and Basidiomycetes.It can produce a variety of active substances through submerged fermentation,among which polysaccharide is the most important one and has a variety of biological activities.At present,the research on A.camphorata polysaccharides mainly focuses on in vitro biological activities such as antioxidant,antiviral,anti-tumor and anti-inflammatory,and there are few studies on the regulation of intestinal flora by polysaccharides from A.camphorata.Long-term use of antibiotics can easily lead to intestinal flora disorder,and then cause symptoms such as diarrhea,inflammation,and immune decline,which damage human health.In this study,the intracellular polysaccharides and exopolysaccharides of A.camphorata by submerged fermentation were characterized first,then the therapeutic effects on diarrhea mice and the regulation of intestinal flora were explored through in vivo experiments.The in vitro experiment verified the effect of polysaccharide from A.camphorata on the growth of lactic acid bacteria.The main contents of this article are as follows:(1)Structural characterization of intracellular and exopolysaccharide polysaccharide from A.camphorata,including monosaccharide composition analysis,molecular weight distribution and infrared spectroscopy analysis.The results showed that exopolysaccharide of A.camphorata(AEPS)was mainly composed of five monosaccharides of galactose,glucose,mannose,glucuronic acid and galacturonic acid,and with contents of 7.84%,84.73%,5.27%,0.76%and 7.84%,respectively;Intracellular polysaccharide of A.camphorata(AIPS)is mainly composed of four monosaccharides of galactose,glucose,mannose and galactosamine,and with contents of 28.52%,55.31%,14.34%and 1.83%,respectively;the average molecular weight of AEPS(MW)is 4.16×105 Da,and the MW of AIPS is 3.52×106 Da;FT-IR spectrum analysis results show that AEPS is mainly ?-glycosidic structure;AIPS has amide structure and contains(-C?C-H),(C-O)functional groups.In vitro simulated digestion experiments show that both AEPS and AIPS have good resistance to ?-amylase and simulated gastric juice digestion ability.(2)The effects of different doses of lincomycin hydrochloride(LIH)by gavage on the state of mice were investigated.It was found that LIH of 3.7 g/kg(mouse body weight)by gavage had adverse effects on hair removal,diarrhea and activity of mice,and the recovery speed after withdrawal was moderate.It was suitable for constructing a mouse model of antibiotic related diarrhea,and then AEPS and AIPS were administered to treat and regulate the intestinal flora of mice.The results showed that AEPS can significantly increase the relative abundance of some beneficial microorganisms(such as Lactobacillus)in the intestinal flora of mice,and significantly reduce the relative abundance of some harmful microorganisms(such as Enterococcus,Staphylococcus).The regulation effect is significantly better than AIPS.(3)Further optimization of the intragastric dose of AEPS found that the intragastric administration of 0.25 g/kg(mice body weight)of AEPS could significantly increase the relative abundance of probiotics(such as Lactobacillus,Roseburia,Ligilactobacillus,Lachnospiraceae_NK4A136_group)in the intestinal flora of mice,significantly reduced the relative abundance of harmful microorganisms(such as Enterococcus,Shigella),and stabilized the relative abundance of Akkermansia in the mouse gut;ultimately restored and maintained the balance of the gut microbiota structure in mice,alleviating the effect of LIH on microbiota.Adverse symptoms such as diarrhea,inflammation,and weight loss caused by rats.(4)The effect of polysaccharide from A.camphorata on the growth of lactic acid bacteria was further verified by in vitro experiments.The results showed that the appropriate concentrations of AIPS and AEPS could significantly promote the growth of four lactic acid bacteria(Lactobacillus casei,Lactobacillus plantarum,Lactobacillus rhamnosus LV-1 strain and Lactobacillus rhamnosus LGG strain).What's more,The promoting effect is not due to the provision of monosaccharide nutrients,but it is likely that one or several polysaccharide components of the polysaccharides.