The Location And Expression Of TFF3in The Intestinal Tract Of Weaning Diarrhea Piglets And The Effect In Testinal Mucous Epithelial Cells

TFF3(Trefoil factor family3) is a mature peptide composed of59AAs. TFF3are known to be expressed in intestinal epithelial cells of various organisms and are often co-secreted with mucins by mucus-producing. TFF3is secreted as a monomer or dimer. TFF3provide protective and reparative effects by enhancing epithelial integrity and promoting mucosal restitution. The expression of TFF3in tissue specificity can be changed in disease condition. Mucosal damage can induced the expression of TFF3. Weaning is one of the most stressful events a pig encounters in commercial swine production. At this time, the newly weaned pig encounters psychosocial and physical stressors including maternal and littermate separation, abrupt changes in diet and environment, and comingling and establishment of social hierarchy with unfamiliar pigs from different litters which can initiation the Early-weaning syndrome. Intestinal epithelial cells are important part of the intestinal barrier. There are many research proof that TFF3is playing a important function in intestinal disease. These processes are of central physiological relevance during the postnatal intestinal development and are strongly influenced during the weaning period. The study was established in the model of diarrhea in piglets. Intestinal TFF3protein expression was evaluated in post-weaning diarrhea piglet. Bacterial endotoxins (lipopolysaccharides (LPS)) are the major outer surface membrane components present in almost all Gram-negative bacteria and act as extremely strong stimulators of innate immunity and inflammation. The expression of mucin and cytokine can be regulated by LPS. This experiment was established in the success of intestinal mucous epithelial cells (IMEC) cultured. Evaluate the effect of the protect the cell not injured from LPS by rhTFF3. The expression of TFF3gene in intestinal epithelial cells would be understood which were treated by different density LPS and quantitative recombinated mutant of human intestinal trefoil Factor-3(rhTFF3).The investigative object were24Duroc piglets (25-28days aged and weighing10±0.3kg),12piglets were infected with Enterotoxigenic E.coli by oral.9piglets with diarrhea were selected as a diarrhea groups, and other9healthy piglets as control group. The piglets were abataged and the samples were collected in the diarrhea lth,3th and5th days in test group and control group (n=3). Immunohistochemisty was used to observe expression difference of TFF3protein in intestinal tissues of piglets. The Image-pro plus of image analysis software was used to detect the positive expression rate of TFF3protein, and the positive expression was the cytoplasm decoration to be yellow. The expression of TFF3protein was indicated by mean density, and results was analysised by SPSS for Windows13.0. The small intestine tissues were separated from newborn piglet, and the intestinal mucous epithelial cells (IMEC) were cultured by using enzymatic digestion. The cells were cultured in the96pore culture plate in the second generation, and the rhTFF3were added after or before the LPS had be added. The MTT measure was used to evaluate the effect of the protect the cell not injured from LPS by rhTFF3. After the cells were overgrow than80percent, the different concentration LPS (1,10,100μg/ml) and rhTFF3(50μg/ml) were added in the medium of cells as the test groups, and the physiological saline was added in the control group. After the cells were acted for24hours, the cDNA of TFF3analysis was carried out using the SYBR green chemistry on a real-time PCR cycler. Furthermore, the analysis the expression of TFF3gene observed which effected by LPS and rhTFF3.We found that TFF3proteins were detected in different amounts in all investigated tissues, and most of them expressed in mucous of intestinal. A increase protein expression levels was observed in all of the intestinal tissues along with the periods of diarrhea, especially in jejunum and ileum. The expressions of TFF3protein were signification compared with control group, and they were detected much more than the control group. It may owe to the function of TFF3which can protect the integrality of intestinal mucous barricade in the post-weaning diarrhea. The TFF3protein was detected in swine intestinal mucous epithelial cell. The injury of LPS can be prevented by rhTFF3in IMEC (p<0.05), and there were different effect in different group which was added different concentration of LPS (p<0.05). It may owe to the different injury by different concentration. Compare with control group. The inhibition expression of TFF3can be detect in light concentration (1,10μg/ml) LPS group, and the higher concentration (100μg/ml) LPS was associated with a distinct response of increased TFF3expression. The expression of TFF3was lower than the control group when the rhTFF3were existence yet.