| Many kinds of botanlic polysaccharides are separated and extracted by modern instruments,and the effect of promoting the growth of animals and regulating the immunity of the organism is shown in the animal.At present,the experimental research on polysaccharides mainly focused on the effect test of single polysaccharides,but the comparison of the effects between single polysaccharides and the combined use of several polysaccharides on the intestinal environment of weaned piglets are rarely reported.This study was conducted to study the effects of Astragalus polysaccharides,Achyranthes bidentata polysaccharides,and Atractylodes macrocephala polysaccharides on the performance and immunity of weaned piglets.The aim was to provide theoretical basis and practical guidance for the popularization and application of high quality piglet diets.The experiment used boiled alcohol precipitation method to extract polysaccharides of Astragalus polysaccharides,Atractylodes macrocephala polysaccharides and Achyranthes bidentata polysaccharides.The contents of these three polysaccharides were determined by the indoxyl sulfate method,and its antibacterial effect in vitro was studied.256 two-day-old crossbred weaned piglets were randomly divided into 8 groups with 4 replicates and 8 pigs each replicate.Three factors and two levels of experimental design were adopted.The amount of Astragalus polysaccharides in the diet was 0 and 800 mg/kg,the amount of Atractylodes macrocephala polysaccharides was 0 and 800 mg/kg,and the amount of achyranthes polysaccharide added was 0 and 800 mg/kg.28 d.The effects of botanlic polysaccharides on the performance of weaned piglets,blood biochemical parameters,intestinal environment,intestinal mucosal immunity,intestinal barriers and related transport carriers were investigated.Three factors and two levels of experimental design were used to test the effects of different plant polysaccharides on intestinal epithelial membrane permeability,glucose transporter,tight junction factor,cell development-related factors,and NF-κB-related factor expression.The main results are as follows:(1)Extraction and antibacterial effect of plant polysaccharidesThe contents of Astragalus polysaccharides,Atractylodes macrocephala polysaccharides and Achyranthes bidentata polysaccharides were 60.3%,60.1% and 65.3%,respectively.Astragalus polysaccharides,Atractylodes macrocephalae polysaccharides andAchyranthes bidentata polysaccharides have certain antibacterial effects on Escherichia coli,Salmonella and Staphylococcus aureus.The antibacterial effect of Astragalus polysaccharides on various bacteria is Staphylococcus aureus>Bacillussubtilis> Salmonella>E.coli.The antibacterial effect of polysaccharides of Atractylodes macrocephala on various bacteria was Staphylococcus aureus>E.coli>Salmonella>Bacillus subtilis.The antibacterial activity of Achyranthes bidentata polysaccharides has a dose-effect relationship,and the relationship of antibacterial activity is: Staphylococcus aureus>Bacillus subtilis>Escherichia coli>Salmonella.(2)Effects of plant polysaccharides on growth performance and blood biochemical parameters of weaned pigletsFeeding astragalus polysaccharide,Achyranthes bidentis polysaccharide,or polysaccharide from Atractylodes macrocephala Koidz in the diets of weaned piglets alone or in combination increased the daily weight gain of weaned piglets(P>0.05).Combined use of the three polysaccharides significantly increased the number of weaned piglets.Weight gain(P<0.05),there was no significant difference in feed conversion between the groups(P>0.05).The addition of astragalus polysaccharide,Atractylodes macrocephala polysaccharide,and Achyranthes bidentis polysaccharide in diets of weanling piglets can significantly reduce the diarrhea rate of weaned piglets(P<0.05),and there is a certain interaction between polysaccharides.All three polysaccharides can reduce the content of glucose,urea nitrogen,total cholesterol,and triglyceride in the blood of weaned piglets and reduce the activity of AST and ALT in the blood.The effect of the three polysaccharides on the glucose in the blood has a synergistic effect.Polysaccharide can significantly increase the content of total protein and high density lipoprotein in serum.The combination of three polysaccharides has a significant effect.(3)Effects of plant polysaccharides on intestinal environment in weaned pigletsPolysaccharides can significantly increase mucosal villus height in duodenum,jejunum and ileum of weaned piglets,increase flock ratio(P<0.05),and effect of three polysaccharides combined polysaccharide group added weaning piglet V/C ratio of ileum was significantly increased(P< 0.05)There was no significant difference between the polysaccharides in each single addition and the V/C ratio was significantly increased compared to the two additions and single additions for the three polysaccharide additions.The pH values ??of the ileum and rectum of weaned piglet fed with single or combined polysaccharide group had no significant change(P>0.05),the pH value of cecal and colon contents was significantly decreased(P<0.05),and the interaction of three polysaccharides added to the cecum contents had interactions.effect.The contents of total volatile fatty acids in the cecal contents of weanling piglets fed with single or combined addition of polysaccharides increased significantly(P<0.05),and the effect of the three combinations was significantly higher than that of the polysaccharide alone group(P<0.05).Addition of plant polysaccharides alone or in combination in weaned piglets diets significantly increased the number of lactic acid bacteria and bifidobacteria in the intestine(P<0.05).The combination of polysaccharides in the two-combined group compared with the single-added group of lactic acid bacteria and bifidobacteria in the intestine The number had an increasing trend,but the difference was not significant(P>0.05).The addition of polysaccharides alone or in combination could significantly reduce the number of E.coli in the intestine(P<0.05).(4)Effects of plant polysaccharides on immune performance and antioxidation of weaned pigletsWhether a single polysaccharide,two polysaccharides,or three polysaccharides combined can significantly increase the serum total antioxidant capacity of weaned piglets(P<0.05),polysaccharides can increase the serum GSH-Px content in weanling piglets,and significantly reduce serum in weaned piglets.MDA content(P<0.05),the combination of three polysaccharides has a higher effect than one polysaccharide.Astragalus polysaccharides,Atractylodes macrocephala polysaccharides and Achyranthes bidentata polysaccharides can significantly increase the serum IgA content in weaned piglets,and the combined effect of the three polysaccharides was significantly higher than that of single polysaccharide(P<0.05).The addition of three polysaccharides alone or in combination had no significant effect on the serum IgM content in weaned piglets,and the single polysaccharide had no significant effect on the serum IgG content in weaned piglets.The single or combined use of the three polysaccharides increased the levels of IL-2,IL-1β,TNF-α and IFN-γ in serum of weaned piglets,and the single polysaccharide had no significant effect on serum IL-6 levels in weaned piglets.Both polysaccharides combined with two or three polysaccharides significantly increased the serum IL-6 level(P<0.05).Astragalus polysaccharides,Atractylodes macrocephalae polysaccharides and Achyranthes bidentata polysaccharides can significantly increase SIgA content in intestinal mucosa of weaned piglets(P<0.05),and the combination effects of the three polysaccharides are the most significant.Combined use of the three polysaccharides can significantly increase the expression of IL-2,IL-1β,IL-6,TNF-α and IFN-γ in the mesenteric lymph nodes and intestinal mucosa of weaned piglets.The combination of the two polysaccharides can significantly increase the mesentery.The expression levels of IL-2,TNF-α and IFN-γ in lymph nodes and jejunum mucosa(P<0.05)had no significant effect on the expression of IL-6 and IL-1β genes in mesenteric lymph nodes and intestinal mucosa.Single added polysaccharide had no significant effect on the expression of IL-2,IL-1β and IL-6 in mesenteric lymph nodes and intestinal mucosa of weaned piglets,and significantly increased the expression of TNF-α and IFN-γ genes(P<0.05).(5)Effect of plant polysaccharides on intestinal barrier in weaned pigletsThe activity of DAO,the content of ET and the content of D-lactose in the serum of weaned piglets from the plant polysaccharide group were significantly decreased,and there was a synergistic effect with the increase of polysaccharide combinations.Plant polysaccharides can significantly increase the gene expression of Occludin,claudin and ZO-1 in the intestinal mucosa of weaned piglets.The single addition of Astragalus polysaccharides,Atractylodes polysaccharide and Achyranthes bidentata polysaccharides significantly increased the relative expression of mRNA(P<0.05)in the intestinal SGLT1 of weaned piglets,and the relative expression of GLUT2 mRNA increased,but the difference was not significant(P>0.05).Any two combinations of three kinds of polysaccharides or combined use of three polysaccharides could significantly increase the relative expression of SGLT1 and GLUT2 mRNA in weaned piglets(P<0.05).(6)Molecular mechanism of plant polysaccharides on protective function of intestinal epithelial barrier in pigletsThe minimum concentration of 60 mg / L of Atractylodes macrocephala Koidz polysaccharides,Astragalus polysaccharides and Achyranthes bidentata polysaccharides on the cell finger value of IPEC-J2 over 35 h was significantly different from that of the control group(P<0.01),and the cell transmembrane resistance(TEER value)had an increasing trend after the addition of the three polysaccharides.The addition of Astragalus polysaccharidealone or combination of three polysaccharides significantly reduced the activity of IPEC-J2 alkaline phosphatase and maltase under LPS(P<0.01).Single Astragalus polysaccharides,Atractylodes macrocephalae polysaccharides and Achyranthis polysaccharides significantly increased the relative expression of SGLT1 and GLUT2 mRNAs in IPEC-J2 cells treated with LPS(P<0.05),while the combination of three polysaccharides significantly increased the relative mRNA expression of SGLT1 and GLUT2(P<0.01).(3)At the level of gene transcription,the expression level of ZO-1 and Occludin mRNA could be improved by either single or combined polysaccharide group(P<0.05),while the combination of Astragalus polysaccharides with Atractylodes polysaccharides or three polysaccharides was significantly increased in the expression of ZO-1 and Occludin protein(P< 0.01).Compared with the control group,the single or combined use of astragalus polysaccharides,atractylodes macrocephalae polysaccharides and achyranthes bidentata polysaccharides could not significantly increase the expression of PCNA and TGF-β in IPEC-J2 cells(P>0.05).Adding Astragalus polysaccharides,Atractylodes macrocephalae polysaccharides and Achyranthes bidentata polysaccharides could significantly increase the mRNA and protein expression levels of NF-κB-related factors TLR4 and MyD88(P<0.01),Astragalus polysaccharides alone significantly increased the expression of NF-κB protein(P<0.01).Therefore,the extracts of astragalus polysaccharides,achyranthes bidentata polysaccharides and polysaccharides from Atractylodes macrocephala Koidz(alone,two,or all three)can reducing the permeability of IPEC-J2 cell membrane after LPS treatment by regulating the expression of SGLT1,GLUT2,Occludin and ZO-1 of the pig small intestinal glucose transporter,in order to maintain the damage and repair of IPEC-J2 cells,and this repair mechanism mainly relied on the activation of TLR/NF-κB signaling pathway.In this paper,we find that plant polysaccharides can improve the production performance of weaned piglets through the research route of "growth performance-intestinal environment-intestinal mucosal immunity and antioxidant performance-intestinal physical barrier-in vitro environment simulation".Adjusting endocrine disorders caused by weaning stress and intestinal damage caused by dimmune decline and intestinal permeability declinedue. |
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