Effects Of ETEC With Gastrointestinal Tight Junction Protein ZO-1 On Early Weanling And Diarrhea Pigs

The early Weaning technology is commonly concerned and used in tensive and large-scale domestic pig-production in advanced technologies. Before and after the piglets in the gastrointestinal tract microflora has changed greatly. Currently many studies suggest that early weaning and diarrhea piglet resulting from the clinical symptoms is mainly due to the organization and function in young piglets digestive tract development has not yet fully matured, weaned piglets of Stress caused gastrointestinal disorders, stomach of piglets, intestinal barrier damaged, resulting in piglets "weaning syndrome" occured. The results show that:the integrity of intestinal mucosa in early weaned piglets have different syndrome critical. Intestinal mucosal barrier, including a complete intestinal epithelial cells and cells form the mechanical connection between the barrier posed by the normal intestinal microbial flora of the biological barrier and intestinal-associated lymphoid tissue and the secretion of antibodies by the immune barrier posed. Tight junctions between intestinal epithelial cells is the main connection to the maintenance of epithelial cell polarity, regulating the permeability of the intestinal barrier to prevent bacteria, endotoxin and toxic macromolecules into the body plays an important role. ZO-1 is the important organizational component of tight junctions, and its expression increases the permeability of the organization can be adjusted. Therefore, the level of detection in molecular biology ZO-1 changes in the mechanism of diarrhea in weaned piglets of great significance, but also decrease the incidence of post-weaning syndrome.Objective:To establish the PCR reaction system, which can simultaneously detect LT, ST-â… and ST-â…¡three kinds of E. coli virulence factors. Detection of the tight junction protein ZO-1 in weaned piglets with ETEC diarrhea of gastrointestinal tissue distribution and expression level and significance;Methods:The experimental ETEC scored 20 days of old piglets were taken 1d,3d, 5d,7d of the gastric mucosa, ileum, colon, semi-quantitative PCR method to explore reaction conditions suitable for real-time quantitative PCR was used to detect gastrointestinal tight junction protein ZO-1 and the internal reference gene GAPDH in mRNA abundance; Using the method of Western blotting to detecte tight junction protein gastrointestinal protein ZO-1 expression.Results:Through this exprement we can successfully establish the three types of ETEC toxin virulence factors LT, ST-â… and ST-â…¡in the PCR detection The real time quantitative PCR showed that ZO-1 mRNA expression level of correlation with tissue differentiation, as the severity of diarrhea in weaned piglets, ZO-1 expression rate of decline. Gastrointestinal tight junction protein ZO-1 expression in three tissues can be can be seen in scoring after ETEC 5d,7d expression differences are very clear expression of time with diarrhea gradually decreasing; The results of Western blotting showed that ZO-1 protein expression decreased the expression level was significantly lower than internal reference gene expression.Analysis:In this experiment we successfully defence the function of control mechanisms to promote the field of basic scientific theory level of the clinical practice and the weaning to provide the technological research about production platforms, for nourishment about regulatory structures and functions of the bowel provide theoretical basis for the prevention and early weaning syndrome "about", and enhance survival rate hewed out a new ideas and promote the beneficiate and margin call.