Effects Of Mongolian Veterinary Medicine Babu-7 And Its Effective Ingredient Compound On Intestinal TLR4/NF-κB Signaling Pathway Of Diarrhea Calves

Diarrhea is one of the most serious diseases of calves,and its damage to calves and economic losses are huge.Previous studies have shown that Mongolian medicine Babu-7and its effective ingredient compound can prevent calf diarrhea without toxic side effects and drug resistance.Therefore,in this study,E.coli O1 diarrhea calves were used as the model to study the effects of Mongolian medicine Babu-7 and its effective composition compound（BSP and BSEIC）on intestinal TLR4 signaling pathway and NLRP3inflammasome of diarrhea calves,providing a theoretical basis for further study of the efficacy and mechanism of Mongolian medicine Babu-7 and its effective composition compound in the treatment of diarrhea calves,In order to find the natural antibacterial drugs to inhibit pathogenic microorganisms instead of antibiotics for scientific basis.The experimental design is as follows:21 newborn Holstein calves with similar body condition were randomly divided into 7 groups:healthy group（NC）,diarrhea group（EC）,ciprofloxacin group（CIP）and Mongolian Veterinary Medicine Babu-7 compound medium dose group（BSP-M）,Mongolian veterinary medicine Babu-7 active ingredient compound low dose group（BSEIC-L）,medium dose group（BSEIC-M）and high dose group（BSEIC-H）,with 3 calves in each group.Calves were dosed twice a day for 7 days.Except the healthy group,the other 6 groups were given 2.50×10¹¹ CFU/m L of pathogenic E.coli O1,100 m L/head on the fifth day.Blood and small intestine tissues were collected 1 h after the last administration on the 7th day.The test methods and results are as follows:Experiment 1:The number of duodenal goblet cells was observed by PAS staining,and the contents of diamine oxidase（DAO）,D-lactic acid（D-LA）in serum and s Ig A and ITF in intestinal tissues were determined by ELISA.The results show that:（1）The contents of DAO and D-LA were significantly increased in EC group,Mongolian medicine Babu-7 and its effective ingredient compound could significantly increase the levels of DAO and D-LA in serum（P<0.05）and were equal to those of the CIP group.（2）The contents of ITF and s Ig A in small intestine of EC group were significantly decreased,Mongolian medicine Babu-7 and its effective ingredient compound could increase ITF and s Ig A content significantly or extremely significantly（P<0.05,P<0.01）.（3）The number of goblet cells in the duodenum of calves in the diarrhea group was significantly reduced,while the number of goblet cells in the BSEIC and BSP groups was significantly increased compared to the diarrhea group.Experiment 2 The contents of catalase（CAT）,glutathione peroxidase（GSH-Px）,malondialdehyde（MDA）,superoxide dismutase（SOD）and total antioxidant capacity（T-AOC）in serum and IL-2,IL-4,IL-10,IFN-γ,TNF-αand TGF-βin intestinal tissue were determined by ELISA.The results showed as follows:（1）Mongolian medicine Babu-7 and its effective ingredient compound could increase the T-AOC,enhance the activities of GSH-Px,CAT and SOD,and decrease the level of MDA in serum of calves（P<0.05）.（2）The contents of ITF and SIg A in small intestine of EC group were significantly decreased,Mongolian medicine Babu-7 and its effective ingredient compound could increase the levels of IL-2,IL-4 and IL-10,and decrease the levels of IFN-γ,TNF-α,TGF-βand IL-1βin different degrees.Experiment 3 The m RNA and protein expression levels of TLR4,My D88,NF-κB,NLRP3 and caspase 1 in small intestine were detected by real-time quantitative PCR（q PCR）and Western Blot.results showed that:The m RNA and protein expression levels of TLR4,My D88,NF-κB,NLRP3 and caspase 1 in EC group were significantly increased（P<0.01）,Mongolian medicine Babu-7 and its effective ingredient compound could significantly reduce the m RNA and protein expression levels of TLR4,My D88,NF-κB,NLRP3 and caspase 1 in small intestine（P<0.05,P<0.01）,and effect of BSEIC-M was the best and better than that of CIP group.Conclusion:Mongolian medicine Babu-7 and its effective ingredient compound could repair intestinal damage caused by pathogenic E.coli O1,protect intestinal mucosal permeability,regulate immune response,and improve intestinal inflammatory damage of diarrhea calves by inhibiting TLR4/NF-κB signaling pathway and NLRP3 inflammatories.