Immunomodulatory Effect Of Eucommia Ulmoides Leaf Extract And Its Application In Broiler Diet

With the characteristics of pure nature,low toxicity,low residue and no resistance,herbal extracts are regarded as the backbone of the farming industry to replace resistance and are increasingly used in the prevention of animal diseases and healthy farming.As a traditional Chinese medicine with medicinal and food properties,Eucommia ulmoides has a long history of application in Chinese medicine and Chinese veterinary medicine.Eucommia ulmoides is mainly used as a medicine for its bark,and the resource utilization rate is low.A large number of studies have shown that Eucommia ulmoides leaves are rich in a variety of natural active ingredients and their pharmacological effects are similar to those of Eucommia bark.Replacing Eucommia bark with Eucommia ulmoides leaves can greatly improve the level of resource utilization of Eucommia ulmoides,which is of great significance to the sustainable development of Eucommia ulmoides and the reduction of costs and efficiency.This study is based on network pharmacology and molecular docking techniques to predict and validate the main active ingredients and related target pathways of Eucommia ulmoides leaves.Exploring the immunomodulatory mechanism of Eucommia ulmoides leaves through in vitro and in vivo assays.The effects of Eucommia ulmoides extracts and ferments on growth performance,immune function and intestinal flora of broiler chickens were investigated through the addition of Eucommia ulmoides to broiler diets,aiming to provide a theoretical basis for the development of feed additive products and farming applications of Eucommia ulmoides.The test results are as follows:1.Network pharmacological analysis and molecular docking verification of immune regulation of Eucommia ulmoides leaves.Screening the TCMSP database to identify the main active ingredients of Eucommia ulmoides leaves,and constructing a PPI network of active ingredient targets of Eucommia ulmoides leaves with immune dysregulation targets;GO enrichment analysis and KEGG pathway analysis of the screened core immunomodulatory targets of Eucommia ulmoides leaves;Reliability of the interaction of the main chemical components of Eucommia ulmoides leaves with key proteins was verified by molecular docking.The results show that the main components of Eucommia ulmoides leaves,such as kaempferol,quercetin and chlorogenic acid,exert immunomodulatory effects by regulating core targets such as the cytokines TNF,IL-6 and IL-1β,and participating in signalling pathways such as MAPK.Quercetin,kaempferol and chlorogenic acid have good binding activity to TNF,IL-6 and IL-1β.2.Immune-enhancing effect of Eucommia ulmoides leaf extract on mouse abdominal macrophages.To determine the effect of Eucommia ulmoides leaf extract on the proliferative capacity and phagocytosis of macrophages and to detect the level of NO and cytokine secretion.The results showed that at concentrations of 2-5 mg·m L^-1,Eucommia ulmoides leaf extract significantly enhanced the proliferative capacity of macrophages and the secretion levels of cytokines TNF-α,IL-6 and IL-1β;At concentrations ranging from 1 to 5 mg·m L^-1,Eucommia ulmoides leaf extract significantly enhanced the phagocytosis of macrophages;At concentrations of 4 mg·m L^-1and 5 mg·m L^-1,Eucommia ulmoides leaf extract significantly promoted the secretion of NO.3.Study on the immunomodulatory mechanism of Eucommia ulmoides leaf extract in immunosuppressed mice.To establish a mouse immunosuppression model using cyclophosphamide and to investigate the immunomodulatory mechanism of Eucommia ulmoides leaf extract in immunosuppressed mice.The results showed that Eucommia ulmoides leaf extract increased body weight and immune organ index in immunosuppressed mice,reduced pathological damage to liver and spleen,inhibited the decline of leukocytes and lymphocytes,improved phagocytosis of macrophages,enhanced ear swelling response,promoted the secretion and expression of inflammatory cytokines,and increased the expression levels of p-p38,p-ERK1/2 and p-JNK.4.Study on the effects of the addition of Eucommia ulmoides leaf extract and fermented matter to the diet on the growth performance,immune function and bone of broiler chickens.The effects on growth performance,immunity and bone function of broiler chickens were investigated through the addition of 1%Eucommia ulmoides leaf extract and 0.6%Eucommia ulmoides leaf ferment to broiler diets.The results showed that both the addition of Eucommia ulmoides leaf extract and fermented matter to the diet improved the growth performance,spleen index,thymus index and bursal index of broilers,reduced the serum cholesterol and triglyceride content of broilers,increased the serum globulin level of broilers,increased the total amino acid and fresh amino acid content of chicken meat,and significantly improved the bone density,increased the area of bone trabeculae and reduced the separation of bone trabeculae in broilers.5.Study on the improvement of intestinal flora of broiler chickens by Eucommia ulmoides leaf extract and ferments.Study of the effects of Eucommia ulmoides leaf extract and ferments on the structural composition of broiler intestinal flora by 16S DNA high-throughput sequencing.The results showed that Eucommia ulmoides leaf extract and fermentation products improved the Alpha diversity of intestinal flora in broilers,and the Beta diversity of intestinal flora in both groups was significantly different from that in the control group.Eucommia ulmoides leaf extract and fermentation significantly increased the ratio of Firmicutes to Bacteroidesl,increased the abundance of Faecalibacterium,and decreased the abundance of Anaerotruncus in the gut microbiota of broilers.To sum up,this study predicted the active components and targets of Eucommia ulmoides leaves through network pharmacology,and verified the immunomodulatory mechanism of Eucommia ulmoides leaves combined with in vivo and in vitro experiments.Through feeding experiments,the effects of Eucommia ulmoides leaf extracts and fermented products in broiler diets and their improvement on intestinal flora were analyzed,which further proved the role of Eucommia ulmoides leaf in promoting broiler growth performance,improving chicken quality and bone performance,and enhancing immunity,and provided theoretical basis for the application of Eucommia ulmoides leaf in livestock and poultry breeding.