Based On The Intestinal Microenvironment, The Immunomodulatory Effects Of Honeysuckle And Lonicera Japonica Polysaccharides Are Discussed

L.JAPONICAE FLOS and L.FLOS have the same effect,both of which are to clear heat and antitoxicant and to disperse wind and heat,they are commonly used in clinical practice.L.JAPONICAE FLOS and L.FLOS also have similar chemical components,in addition to the main active components of organic acids,flavonoids,saponinas.L.JAPONICAE FLOS and L.FLOS also contain some polysaccharides,polysaccharides have certain immunomodulative activity,there is no comparative study on polysaccharide structure and immune regulatory activity of L.JAPONICAE FLOS and L.FLOS.The intestinal microenvironment is closely related to the immune function of the body,so the intestinal microenvironment can be used as a starting point to compare the immunomodulatory activities of L.JAPONICAE FLOS polysaccharides and L.FLOS polysaccharides.In this study,ultrasonic assisted extraction was used to extract polysaccharides from L.JAPONICAE FLOS and L.FLOS Based on single factor experiment,orthogonal experiment design was adopted to optimize the extraction process of polysaccharides.The crude polysaccharides of L.JAPONICAE FLOS and L.FLOS were obtained by water extraction,alcohol precipitation and deproteinization of Savag,which were used to study the immunomodulative activities of polysaccharides.The pure polysaccharides of L.JAPONICAE FLOS and L.FLOS were obtained by DEAE-52 cellulose column chromatography and Sephadex G-100 glucan gel column chromatography,which were used to study the structure of polysaccharides.With crude polysaccharides of L JAPONICAE FLOS and L.FLOS as the research object,using cyclophosphamide immunosuppressive mice model was established,compare each viscera index,serum levels of cytokines in mice,serum immunoglobulin,intestinal mucosal immune globulin,research crude polysaccharides of L.JAPONICAE FLOS and L.FLOS differences of immunosuppressive mice immune regulating activity;The fecal and colonic contents of immunosuppressed mice were used as the research object to compare intestinal flora abundance and short-chain fatty acid levels in colonic contents,and to study the effects of L.JAPONICAE FLOS and L.FLOS crude polysaccharides on intestinal microenvironment.The pure polysaccharides of L.JAPONICAE FLOS and L.FLOS as the research object,The gas chromatography was used to study the monosaccharide composition of polysaccharides.Ultraviolet spectrum was used to study exists in polysaccharide nucleic acid and protein,Infrared spectroscopy was used to study the glycoside bond configuration of polysaccharides and identify the main substituents on the sugar bonds.Nuclear magnetic resonance was used to identify key main substituent on sugar and spectroscopy to determine the glycosidic bond configuration,sugar chain connection position,anomeric carbon configuration,etc.SEM was used to study morphology characteristics of the polysaccharide.The main results are as follows:1.The optimal extraction conditions of ultrasonic assisted extraction of polysaccharide were as follows:ultrasonic time 30 min,ultrasonic temperature 60℃,solid-liquid ratio 1:30;The optimum extraction conditions were used to respectively extract polysaccharides from L.JAPONICAE FLOS and L.FLOS.The extraction rate of polysaccharides was 20.74%from L JAPONICAE FLOS and 21.53%from L.FLOS.The crude polysaccharides of L.JAPONICAE FLOS and L.FLOS were obtained by alcohol precipitation and deproteinization.The yield of crude polysaccharides of L.JAPONICAE FLOS and L.Flos.was 2.16%and 2.96%,and the content of polysaccharides was 32.34%and 34.92%.The crude polysaccharides of L.JAPONICAE FLOS and L.FLOS were purified by column chromatography.The water eluent was collected and lyophilized to obtain the purified polysaccharides of L.JAPONICAE FLOS and L.FLOS.The yield of polysaccharides of L.JAPONICAE FLOS and L.FLOS was 1.15%and 1.41%.The content of purified polysaccharide was 96.31%and 94.15%.In other words,The polysaccharide content of L.FLOS was higher than that of L.JAPONICAE FLOS.The yield and content of crude polysaccharide of L.FLOS were higher than that of L.JAPONICAE FLOS.The yield and content of polysaccharides of L.FLOS were higher than that of L.JAPONICAE FLOS2.Both crude polysaccharide from L.JAPONICAE FLOS and L.FLOS significantly increased thymus index and spleen index in immunosuppressed mice,and the effect of the high-dose group was better than that of the low-dose group.The crude polysaccharide of L JAPONICAE FLOS and L.FLOS significantly promoted the secretion of IL-6 in immunosuppressed mice,while the crude polysaccharide of L.FLOS significantly promoted the secretion of IL-2 and TNF-α in immunosuppressed mice.The crude polysaccharide of L.JAPONICAE FLOS and L.FLOS significantly increased the content of IgG and IgM in immunosuppressed mice,while the crude polysaccharide of L.FLOS significantly increased the content of IgA.The content of SIgA in colon content was significantly increased by L.JAPONICAE FLOS crude polysaccharide.In other words,L.JAPONICAE FLOS and L.FLOS polysaccharides had significant regulatory effects on the immune function of cyclophosphamide immunosuppressed mice,and there was no significant difference between them.3.The activity of L.JAPONICAE FLOS polysaccharides was better than that of L.FLOS polysaccharides to improve the community richness and diversity of intestinal flora in immunosuppressed mice.The results showed that the microbial composition of L.JAPONICAE FLOS polysaccharides group and L.FLOS polysaccharides group had certain similarity,and there were significant differences with the model group;The distribution of intestinal flora of L.JAPONICAE FLOS polysaccharides was closer to that blank control group than that of L.FLOS,which indicated that L.JAPONICAE FLOS polysaccharides could significantly improve the richness and evenness of intestinal flora,improve the diversity and species composition of intestinal flora in immune suppression mice.There was no significant effect of L.FLOS polysaccharides,but there was a certain improvement trend.The contents of propionic acid,acetic acid,n-valic acid and iso-valic acid in colon contents were significantly increased in the high-dose group of L.JAPONICAE FLOS and L.FLOS crude polysaccharides;the n-butyric acid content was significantly increased in each treatment group of L.JAPONICAE FLOS crude polysaccharides,and the isobutyric acid content was significantly increased in the high-dose group of L.FLOS crude polysaccharides.4.L.JAPONICAE FLOS and L.FLOS polysaccharide has the same kinds of monosaccharide composition,both for the arabinose,rhamnose,ribose,xylose,mannose,fructose,galactose and glucose,and high content of L.JAPONICAE FLOS in the monosaccharide composition of polysaccharides in monosaccharide glucose,fructose and mannose,high L.FLOS the monosaccharide composition of polysaccharides content of monosaccharide glucose,rhamnose,mannose,L.JAPONICAE FLOS and L.FLOS polysaccharide monosaccharide composition of monosaccharide types are the same,the relative content of each monosaccharide there exist certain differences;The polysaccharides of L.JAPONICAE FLOS and L.FLOS not contain nucleic acid and protein by ultraviolet spectrum scanning.There was no significant difference in the infrared spectra of L.JAPONICAE FLOS and L.FLOS polysaccharides,so it was speculated that they should have similar functional group composition.There were characteristic absorption peaks of polysaccharides,β-glycoside bond and α-glycoside bond,pyranoid glycoside bond,etc;1H-NMR and 13C-NMR spectra showed that there was no significant difference in polysaccharides structure between L.JAPONICAE FLOS and L.FLOS.,combined with monosaccharide composition and infrared spectroscopy analysis,polysaccharides in L.JAPONICAE FLOS and L.FLOS were mainly composed of a-D-glucose and a certain amount of β-D-fructose;Both L.JAPONICAE FLOS and L.FLOS polysaccharides were generally smooth and close flaky on the surface under the SEM scan.Under high magnification,it was observed that L.FLOS section had multi-level flaky structure and local surface had irregular texture,while no such structure was observed in L.JAPONIC AE FLOS polysaccharides.In other words,the polysaccharides of L.JAPONICAE FLOS and L.FLOS have some differences in structure,but their similarity is higher.In a word,polysaccharides from L.JAPONICAE FLOS and L.FLOS had certain differences in extraction rate,structure and immune regulatory activity,but the overall difference was small and the similarity was high.This study can provide reference for the application of L.JAPONICAE FLOS and L.FLOS polysaccharides and the evaluation of their immunomodulative activity.It can be used for the development of L.JAPONICAE FLOS and L.FLOS polysaccharide health care products.