Effect Of Berberine On Intestinal Inflammation Caused By Salmonella And Its Therapeutic Effect

In the early stage of this study,the effect of berberine(BBR)on intestinal epithelial cell inflammation induced by Salmonella lipopolysaccharide(LPS)was determined through in vitro experiments.Subsequently,the animal model of Salmonella infected chicks was established in vivo to detect the effects of BBR on immune function,intestinal flora,transcriptome and curative effect of chicks,In order to provide new strategies and candidate traditional Chinese medicine for the prevention and treatment of salmonellosis.1.CCK-8 method was used to determine the optimal treatment concentration,time and administration sequence of IPEC-J2 treated with LPS and BBR;Subsequently,the m RNA expressions of IL-1β,IL-6,ZO-1 and Occludin in the four groups were detected by RT-q PCR,the distribution and localization of Occludin and ZO-1 proteins and their fluorescence intensity were detected by immunofluorescence,and the expression levels of occludin and ZO-1 tight junction proteins were detected by Western blot.The results showed that compared with BBR group and LPS group,the mRNA expression levels of IL-6 and IL-1βin BL group decreased significantly(P<0.05),and the m RNA expression levels of ZO-1 and Occludin increased significantly(P<0.05);Immunofluorescence and Western blot showed that the expression of Occludin and ZO-1 protein in BL group increased significantly compared with LPS group(P<0.05).2.One hundred and twenty 1-day-old Sanhuang chickens(male)with similar body weight were randomly divided into 4 groups:control group(C),berberine group(B),Salmonella challenge group(S)and berberine treatment group(BS).The broilers in group B and BS were supplemented with 120 mg/kg berberine(BBR)respectively.Group S and group BS were gavaged continuously for 3 days at the age of 9days 1×10~9CFU/m L Salmonella,group C and group B were gavaged with the same amount of normal saline as the control group.Detection in the four groups toprevent broiler growth performance,immune organ index,physiological and biochemical indexes,immune index and the antioxidant index.The results showed that the thymus index of 21 day old BS group was significantly lower than that of group C(P<0.05),and that of 35 day old S group was significantly lower than that of group C(P<0.05).The concentrations of Ig M and Ig G in group S were significantly lower than those in group B and BS(P<0.05);The concentration of Ig A in group S at 35 days old was significantly lower than that in group C(P<0.05).The total antioxidant capacity of serum in group s at 21 days old was significantly lower than that in group C(P<0.05).The total antioxidant capacity and high density lipoprotein concentration in group B were significantly higher than those in group C(P<0.05).3.Detecting the intestinal morphology and intestinal flora diversity of four groups of chickens.The results showed that Salmonella increased the abundance of Brucella and other pathogenic bacteria,and decreased the abundance of Bacteroides and Lactobacillus.The addition of BBR increased the abundance of probiotics such as Clostridium,Bacteroides and Lactobacillus,and decreased the abundance of pathogenic bacteria such as Streptococcus.4.The molecular mechanism of berberine regulating Salmonella induced enteritis was preliminarily revealed by cecal transcriptome technology.Go analysis results:the differential genes are mainly in biological processes,including T cell selection,B cell receptor signal pathway,complement activation,phagocytosis and humoral immune response.KEGG enrichment analysis results:the differentially expressed genes were significantly enriched to NF-κB,B cell receptor,Ig A intestinal immune network,calcium signaling pathway and other pathways.In conclusion,BBR can down regulate proinflammatory factors IL-6 and IL-1 in vitroβUp regulate the expression of tight junction associated protein and alleviate the inflammatory response of intestinal epithelial cells;In vivo,BBR has a good effect on Salmonella induced enteritis in chicks,and can regulate the immune response level and intestinal flora imbalance of chicks caused by Salmonella.This process is mainly affected by the positive regulation of biological processes,which is similar to NF-κB.B cell receptor pathway,Ig A and intestinal immune network;The results show that berberine has potential clinical value in the treatment of Salmonella enteritis in chickens.