Expression Of ANG4 Gene In Intestinal Tract Of Newborn Piglets

ANG4 is novel antimicrobial protein which contains 144 amino acid residues and it belongs to ribonuclease A family.Ang4 is mainly expressed in intestinal tissues of mammals.It has the activities of antibacterial and angiogenesis which plays an important role in the regulation of the vascular structure and the remodeling and in the immune process of pathogens.Moreover,ANG4 is also involved in intestinal inflammatory reaction.However,most studies on ang4 are based on experimental mice,but the expression of ANG4 in piglet is not clear.This is also the purpose of this study to understand the expression characteristics of ANG4 in the intestinal tract of piglet.To investigate the fundamental expression pattern of ANG4 in intestine of piglets,20 healthy 1 to 5 weeks of newborn piglets' intestinal samples were collected and the proteins were extracted.The expression level of ANG4 in different segments of intestine at different developmental stage were determined by western blot analysis,and the cell types of ANG4 expression were investigated by immunohistochemistry.The main contents of this paper include the following aspects: 1.ANG4 gene expression levels in different segments of the intestinal tract of newborn pigletsIn order to compare ANG4 gene expression levels in newborn piglets at different developmental stages in different segments,20 healthy newborn piglets of 1 to 5 weeks collected and the protein was extracted from different segments of the intestine(duodenum,jejunum,ileum and colon tissue),RIPA strong lysis buffer and protease inhibitors.The protein samples were analyzed,using western blot detect the expression levels of ANG4,results as followings:The change trend of ANG4 is roughly the same in different parts of neonatal piglet intestine,and in the same intestine segment,ang4 level increased gradually with the age.In the first two weeks the increasing trend is not obviously,it is significant from the third to fifth weeks.In addition,the expression of ang4 in ileum and colon was higher in the same period,followed by jejunum,and the lowest expression in duodenum.2.Determine of cell types of ANG4 expression in piglets? intestineIn order to determine the cell types of ANG4 expression in different segments of piglet intestine,the intestine samples from 3 weeks old healthy newborn piglets of different parts of the intestine(duodenum,jejunum,ileum and colon tissue)were collected and analyzed using immunohistochemical streptavidin peroxidase(streptavidin peroxidase SP)method,The results were as followings: The expression of ang4 protein in different parts of the small intestine is mainly concentrated in the small intestinal villi,but also a small amount of expression in the crypt,the main expression cells are globlet cells and Paneth cells.