| The aims of this study was to investigate the effects of two traditional Chinese medicine prescriptions(Xiao Jian Zhong plus-minus compound(XJZ)and Jiansananli-sepis(JSS)powder)on TLR4/My D88/NF-κB colonic inflammation signal pathway,caecum microbiota and nutrient apparent digestibility in piglets.A total of 18 piglets(Large White× Landrace × Duroc),with similar weight and at 40 days of age,were randomly allocated into Control,TCM1 and TCM2 group based on gender(n=6).Control group was supplied with basal diet,while TCM1 and TCM2 groups were respectively supplied with 10 g/kg XJZ prescription and 3 g/kg JSS prescription in basal diet.After a 5-day adaptation period,the formal trial lasted 60 days.The diarrhea rate of piglets was calculated every day based on diarrhea index.The nutrient apparent digestibility and p H-value in the middle of colonic and cecal contents were determined.The expression of inflammatory related genes and proteins in the TLR4/My D88/NF-κB signaling pathway were measured by real-time quantification PCR(RT-q PCR)and Western blot.High throughput sequencing of16S-r RNA was performed using cecum contents.The results of present study as follows:(1)Compared to the Control group,both TCM1 and TCM2 significantly decreased(P<0.01)the diarrhea rate of piglets,but no difference was observed between TCM groups(P>0.05).The p H value of colon and caecum were decreased after TCM intervention.TCM1 decreased the p H value of colon and caecum by 7.85%(P<0.05)and 11.54%(P<0.01)respectively,and TCM2 decreased that by 1.81%(P>0.05)and 8.73%(P<0.01)respectively.(2)Compared to the Control group,TCM1 remarkably elevated the apparent nutrient digestibility of CP(5.04%,P<0.01),CF(6.63%,P<0.01),NDF(12.04%,P<0.01),ADF(17.12%,P<0.01),Ca(6.56%,P<0.01)and P(6.31%,P<0.05),respectively.Meanwhile,TCM2 significantly promote the levels of CP,NDF and ADF by 2.56%(P<0.05),7.10%(P<0.05)and 13.64%(P<0.01),however,the digestibility levels of CF,ADF,Ca and P were not affected(P>0.05)by TCM2.(3)Compared to the Control group,TCM1 treatment reduced the m RNA levels of TLR4 and My D88 respectively(P<0.05).Meanwhile,TCM2 significantly decreased the m RNA expression of My D88(P<0.01)and NF-κB(P<0.05).Moreover,both TCM1 and TCM2 treatments significantly inhibited the protein levels of NF-κB(P<0.05 or P<0.01),p-NF-κB(P<0.01)and p-IκB-α(P<0.05 or P<0.01),but remarkable increased IκB-αprotein level(P>0.05).(4)Compared to the Control group,The gene expressions of IL-6(P<0.01),IL-8(P<0.01)and TNF-α(P<0.05 or P<0.01)were significantly decreased in both TCM1 and TCM2 groups,but the m RNA level of IL-10 was increased significantly(P<0.01).Meanwhile,the m RNA expressions of IL-8 and TNF-α were down-regulated(P<0.05),but the IL-10 level was significant elevated with TCM1 group than that of TCM2 group(P<0.01).(5)Compared to the Control group,in the phylum level,TCM1 remarkable improved the abundance of Actinobacteria and Cyanobacteria(P<0.01),and TCM2 only increased the abundance of Actinobacteria significantly(P<0.01).In the genus level,the abundance of Eubacterium,Bifidobacterium and Streptococcus were significant elevated in both TCM groups(P<0.05 or P<0.01).Meanwhile,only the abundance of Bulleidia was increased in the TCM1 treatment significantly(P<0.01),but the abundance of Turicibacter was reduced in the TCM2 group remarkably(P<0.05).In conclusion,XJZ plus-minus compound and JSS powder decrease diarrhea rate and elevate nutrient apparent digestibility of piglet by reducing the p H value and ameliorating the overall health of intestinal.The possible mechanism is that the traditional Chinese medicine compound relieve intestinal inflammation,directly or indirectly,via inhibiting the TLR4/My D88/NF-κB inflammatory signaling pathway and improving the abundance of beneficial bacteria of gut in piglets. |
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