| PrefaceLong term and large quantity of alcohol drinking cause dysfunction of multi-organs, such as liver, kidney, heart and pancreas, and present some specific clinical manifestations. Some non-specific digestive symptoms such as abdominal distention, anorexia and nausea cannot be explained convincingly by the above dysfunction. It is suggested that ethanol involed in the gastrointestinal mucosal injury, especially the gastric mucosal injury. A lot of animal experiments indicated that short term and high concentration of ethanol caused acute gastric mucosal injury and induced the up-rugulation of some cytokines expression. There were few studies on the gastric injury caused by relatively long term and low concentration of ethanol. In this study, a chronic alcoholic gastric injury model was set up by oral-gastric ethanol administration. The aim was to explore the pathological changes of gastric mucosal and expression of TGF-a in different stages and, which can promote the proliferation and differen tiation of gastric epithelial cells.Materials1. Animals46 female Wister rats, 16 rats in control group and 30 rats in model group.2. Reagent (l)Anhydrous alcohol(2)Mice-anti-rat antibody: TGF-a( 1:100) , Strept-Avidin-Biotin (SABC) test kit and diaminobenzidine(DAB) showing color reagent.3. Instrument(1) Paraffin microtome(2)Light microscope and photography apparatus(3) Micro (fluorescence)-picture analyze systemMethods1. Animal model46 rats were subjected to two groups randomly. There were 16 rats in control group and 30 rats in model group. The model rats were given alcohol at a dose of 40% , 8g kg-1 day-1, three times a day by intragastric perfusion for the first 4 weeks. 50% alcohol 9g kg-1 day-1 , three times a day for the second 4 weeks and 50% alcohol, 10g kg-1 day-1, three times a day for the third 4 weeks. The control group was treated with physiological saline in the same way as model group. 10 , 12,8 rats in model group and 5,5,6 rats in control group were killed respectively at the end of 4 ,8,12 weeks .2. Pathologic examinationThe stomach was sheared along the larger curve and was put plainly on the glass plate after washing, The injury degree was measured with micro-rule under the low power light microscopy. Lesion index was calculated by Guth criterion. The whole stomach was cut par-alleled and laterally every 0. 3 cm, then paraffin blocks were made. The paraffin block was cut continuously with the thickness of 5um, HE stain was made routinely and the pathologic changes were observed under the light microscopy. Histologic index of gastric mucosa was calculated according to Mascuda criterion.3. Detection TGF-a by ABC immunohistochemistryThe paraffin sections were dewaxed to water, then the endogenet-ic enzyme was inactived. Repair the antigen, add normal rabbit serum , then add the first antibody TGF-a ( 1:100) , stay overnight at 4C and add the second antibody 20 -37C for twenty minutes. ABC compound was added for twenty minutes under 20 -31C. All steps were completely washed by 0. 01M PBS. DAB showed color. The sections were dyed again slighdy by hematoxylin and observed under the light microscope. The cells containing light brown particulate in cell plasm were positive cells. Utilize the micro fluorescence picture analysis system to analyze the integrated intensity of immunohistochemistry of TGF-a in selective field of vision.4. Statistic analysisThe data was analyzed by SPSS10. 0 software , using ANOVA test.Results1. General conditionThe general condition of rats in model group comparing with that in control group was bad. The fur of rats in model group was lackluster. They were irritable and body weight were lost(P <0.01) .2. Stomach pathologic changesIn control group, stomach tissue showed the normal gastric rauco-sa. At the end of 4 weeks, there are dot erosion, hyperemia and edema , neutrophil infiltrated the mucosa. At the end of 8 weeks, gastric mucosa showed multi-erosion, chief cell and parietal cell s...
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