| Objective:1.Evaluation the animal models repeatability of TNBS-induced PI-IBS visceral hypersensitivity;2. The second objective of the study was to investigate the PI-IBS visceral hypersensitivity rat colon pain perception and the shape and the numbers of MC, tryptase and substance P expression and regulation, and to explore the mechanism of acupuncture clinical efficacy in patients with IBS symptoms of abdominal pain or discomfort.Methods:1.30 female Wistar rats were randomly divided into model group (n= 15) and control group (n= 15), the model rats anus about 8.0cm, disposable be TNBS/50% ethanol mixture 0.8 ml (5mgTNBS dissolved in 50% ethanol) enema slowly injected control group,0.8ml saline enema slowly, after the model randomly selected two groups of two rats in each groups,at 7 and 28 days trip intestinal pathology testing, and in the 28 days of observation visceral feeling and form groups and number of colonic mast cells.2. According to a 2:1 ratio,45 female Wistar rats were randomly divided into control group (n= 15), and the preparation of group model (n= 30), modeling completed 30 rats were randomly divided into EA group (n= 15) and model group (n= 15). EA group with EA bilateral Tianshu intervention 14 days, through the pain perception threshold assessment, HE staining, toluidine blue and immunohistochemical methods were observed in three groups of rats visceral sensation, colon pathology specimens, mast cell morphology and number tryptase and substance P expression.Results:Experiment one:By observing the visceral sensory evaluation, detection of colon and colon histopathology MC counting of model group and the control group of rats in the first 28 days after modeling changes, compared with the control group, model group rats intestine mucosal epithelial integrity, epithelial lamina propria few inflammatory cell infiltration, pain perception threshold and maximum capacity sensory thresholds were increased, increasing the number of mast cells, consistent with the clinical manifestations PI-IBS.Experiment two:1.Visceral sensory evaluation:EA group compared to the untreated group, causing the abdomen to lift the balloon capacity threshold increased significantly, and the difference was statistically significant (P<0.05).2.Toluidine blue:EA group mast cells seen under a microscope morphology compared with the model set of rules, oval, surrounded by little purple piece flocculent, the degree of particle removal is not obvious, the number of mast cells in the colon specimens compared with EA group model group, the difference was statistically significant (P<0.05).3. Immunohistochemical results:1) The abdominal pain of the model rats raised thresholds were presented with tryptase and total protein expression of SP material negative correlation (tryptase: r=-0.808, P= 0.015; substance P:r=-0.915, P= 0.001); overall expression levels of tryptase positive colon protein expression correlated with the amount of material was SP (r= 0.879, P= 0.04); 2) EA group tryptase weak staining, small amounts expressed in between intestinal mucosa lamina propria mesenchymal cells, compared with the model group, IOD value difference was statistically significant (P<0.05); 3) EA group SP immunoreactive staining is weak, little expression in the cell membrane and submucosal glands lower, IOD value lower than the model group, the difference was statistically significant (P<0.05).Conclusions:1.About 8cm from the anal margin at one time to be TNBS/50% ethanol mixture 0.8ml (5mgTNBS dissolved in 50% ethanol) slow enema preparation PI-IBS is an ideal animal model program, the model successfully prepared the first 28 days, the performance of rats PI-IBS with a variety of clinical features similar changes, preparation TNBS-induced PI-IBS visceral hypersensitivity model method has good repeatability.2. EA can reduce the number of colon tissue model MC degranulation status and reduce tryptase and protein SP, improve visceral pain perception threshold, and visceral pain relieve allergies. Indicate that acupuncture and moxibustion treatment of patients with IBS symptoms such as abdominal pain or discomfort may be through improving the mechanism of MC activation state and reduce the expression of substance P. |
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