| Soybean is rich in nutrition,not only must have high amino acid content,but also the amino acid is relatively balanced,so soybean and tempeh have been high-quality plant protein sources for livestock and poultry,but anti-nutritional factors such as α-conglycinin,β-conglycinin,Glycinin,etc.can activate the neutrophil to generate an innate immune response to cause intestinal allergic symptoms in young animals such as piglets and give livestock The aquaculture industry has caused economic losses.To address the harm of anti-nutritional factors such as conglycinin to the health of young animals,the detailed mechanism of α,β-conglycinin damage to the intestinal mucosa of young animals must first be clarified.As the body’s first line of defense,neutrophils in the intestine can be degraded by response reactions such as phagocytosis and degranulation after contact with the antigen.In recent years,it has been found that neutrophils can form neutrophils after activation.The extracellular structures of extracellular traps(NETs)are involved in the immune response of the antigen.Studies have shown that NETs is a double-edged sword.In addition to killing pathogens,it also participates in the pathophysiology of many diseases.When NETs are formed too much or cannot be degraded in time,they may cause immune damage to their cells and tissues.A large number of research reports have confirmed that soy antigen proteins can cause an immune response in the intestinal system.However,the effects of antigenic proteins such as α,β-conglycinin on NETs produced by neutrophils and the possible consequences have not been reported.In this paper,in vitro experiments were conducted to observe and verify the effects of alpha-alpha-soy globulin and alpha-alpha-soy globulin on the neutrophils of mice.Furthermore,through in vivo experiments,β-conglycinin was selected for intragastric administration of mice,and a β-conglycinin mouse diarrhea allergy animal model was constructed.The aim is to provide preliminary research basis and theoretical basis for effectively increasing the utilization value of tempeh in production practices and reducing the incidence of intestinal diseases in young animals.Firstly,neutrophils were obtained from the blood of mice by density gradient centrifugation.The effects of α and β-conglycinin on the formation of NETs and their underlying mechanisms were investigated using laser confocal microscope and western blotting.The results show that α,β-conglycinin can induce the release of NETs with DNA as the backbone,and fix protein particles such as elastase and myeloperoxidase.The mechanism may depend on the increase of reactive oxygen species production,activation of MAPK signal transduction pathway ERK1 / 2 and p38 expression.Secondly,the animal model of mice diarrhea allergy was established by using estrogen-associated soybean globulin,and histopathological sections were prepared.The nucleic acid content in serum was tested by Pico~? Green,and the serum IgE level was detected by ELISA,so as to preliminarily evaluate the effect of estrogen-associated soybean globulin on diarrhea allergy in mice.Histopathological results showed that the small intestinal villous epithelial cells of the β-conglycinin group showed local focal necrosis,and the lamina propria of the small intestine showed infiltration of inflammatory cells such as lymphocytes and neutrophils,and DNase Ⅰ pretreatment.Can reduce the histopathological damage caused by β-conglycinin;nucleic acid results show that β-conglycinin significantly increases the content of nucleic acid in serum,and the generated nucleic acid can be degraded by DNase Ⅰ treatment,indicating that the serum of diarrhea mice NETs can be detected in ELISA;ELISA results showed that β-conglycinin caused a significant increase in IgE levels in mouse serum,and DNase I pretreatment could reduce the increase in IgE levels caused by β-conglycinin.The above results indicate that the level of NETs in the intestinal immune system is closely related to the degree of intestinal tissue damage.Based on the above studies,this paper has initially confirmed that α,β-conglycinin induces neutrophils to produce NETs in mice by activating ROS production and MAPK signal transduction pathway ERK1 / 2 and p38 protein expression.A mouse allergy model was established to investigate the effect of NETs on the intestinal innate immune system caused by β-conglycinin,so as to provide theoretical basis and guidance for the prevention and treatment of intestinal allergy in young animals. |
