A Study Of Esophageal Mucosal Cells Apoptosis-related Factor With Gastroesophageal Reflux(GER)

Objective: This study through set up the Reflux Esophagitis animal model (a simple acid reflux, mixed reflux) and sham animal model, to observe the survival and esophageal specimens show the naked eye, detect Esophageal histology specimens,and detecting TNF-α,Caspease3,Survivin by immunohistochemistry. Discussing the role of apoptosis-related factor in the reflux of lower esophageal mucosal cell,the final purpose is to explore directory theory for curing gastric oesophagea reflex disease.Methods: 200 male SD rats were randomly divided into 3 groups: sham group (Group A n=40), Simple acid reflux group (Group B n=80), Mixed reflux group(Group C n=80). Each group was also randomly divided into 20,24,28,32weeks groups in term of the days after surgery(n=8).Rats were killed in the 20, 24, 28,32 weeks after surgery. In obstructed Lower esophageal tissues, histological changes were observed by HE stain, TNF-α,Caspease3,Survivin were examined by immunohistochemical method. The results were analyzed semi-quantitatively with the pathological image analysis system. All the data were analyzed by SPSS13.0statistics software, P<0.05 was considered to have statistical significance. Results: The body weight of the rats marked decline after the experimental, significantly difference compared with group A(P<0.01). After 20 days, most rats weight returned to preoperative levels,However, compared with the sham group slower growth. HE staining: all levels of structural integrity of group A of rats esophageal mucosa thickness and moderate mucosal surface can be seen less keratinized. Rat model of group B with time, gradually thickened mucous layer, basal cell layer and the spine increase in the number of cell layer cells, keratinized layer gradually thickened, we can see mucous layer of surface erosion appears small after 32 weeks; lesions of group C than the same period of the performance of group B significantly heavier, with the exception of the occurrence of such lesions,the part of rat esophageal mucosa partial basal cell layer hyperplasia obvious in 24 weeks, some mucosal tissue showed outward nipple-like protrusions, spines cell layer levels increased significantly, granular layer cells appear to increase, cell Keratocyst situation became clear; model group with time, the scope of mucosal hyperplasia significantly increasing, from the esophagus and stomach and duodenal anastomotic surgical site to the progressive development of the middle and even upper esophagus, papilla like protrusions are also markedly increased. Immunohistochemistry:In the group A at different time points, TNF-αprotein in the cell layer can be seen spine weak positive expression. group B, group C of TNF-αprotein expression was gradually increasing trend; With the same period in group B compared to 28,32-week rat model of group C TNF-αprotein expression increased significantly (P <0.05). Caspease-3 in normal control group of basal cell layer and cell layer cells in the spine has basic positive expression, group B, group C with the time, the expression of the rate of positive cells gradually increased, Positive cells also increased from the basal cell layer to the spine as well as the whole cell layer, group C with the same period in rat model compared with group B to the first week of 28,32 mucosal expression of the rate of positive cells was significantly increased. Normal control group at each time point, Survivin protein expression were not found, Group B animals at the time of 20 weeks of esophageal mucosal layer no nuclei positive expression of Survivin protein, 28,32 weeks the number of positive cells markedly increased the expression, positive expression of parts of lower gray value; group C at each time point in rat model of positive cells in gray value and number of positive cells significantly higher than A.Conclusion: Single acid reflux, mixed reflux surgery can cause rat model of esophageal epithelial apoptosis changes, Mixed reflux surgery acid reflux than the single effect of surgery become more evident, Mixed reflux after surgery 24 Guangyong endoscopic mucosa may appear on piquiri spine at the end of cell layer and cell layer cells in the level of increase, atypical hyperplasia of the region to increase apoptosis. Immunohistochemistry shows that the model group, TNF-αexpression increases, Caspease-3 upregulation, Survivin protein expression increased, Prompt acid reflux, duodenal juice reflux stimulated by the impact of a series of apoptosis-related gene protein expression to cause esophageal mucosal epithelial cell apoptosis.