Effects Of Rubbing Method On Mast Cell Activation In Visceral Hypersensitive Rats With Irritable Bowel Syndrome

Objecttive1.Evaluate the certainty and reproducibility of the IBS visceral highly sensitive rat model reproduced by chronic mother-infant separation method combined with chronic mild stress method through experiments.2.The effects of rubbing method on the visceral sensitivity and behavior of highly visceral sensitive IBS rats were observed through experiments.3.Observation of the effect of rubbing method on mast cells in highly visceral sensitive IBS rats was carried out through experiments.The morphology and number of mast cells and the degree of damage to colon tissue after mast cell activation were explored in order to clarify the mechanism of action of rubbing method on IBS.Methods1.After selecting two pregnant rats and waiting for their normal offspring,the offspring are randomly divided into a model group(n=10)and a normal group(n=10).In the model group,the offspring were raised in the same cage as the mother rats on the first day of birth,Since day 2 the mother and the offspring were separated for 3 hours a day,No intervention measures were taken during the separation,and the mother and child mice could not see each other,and they were kept normally.From the 22 nd day,the mother and child rats were completely separated and kept normally.Starting from 41 days,daily fasting and water fasting are alternated.The normal rats are kept normally for 21 days,starting from the 22 nd day,and then separated and reared normally.After 2 months of age,the model building was completed,and the model was checked with the AWR score、general conditions of rats、water avoidance experiment.2.3 pregnant rats were selected,and after their normal birth,the rats were randomly divided into a model group(n=20)and a normal group(n=10).After successful modeling,the model group will be randomly divided into experimental group(n = 10)and The model group(n = 10)were tested for rat behavior(ie open field test,AWR score).The experimental group adopted rubbing method intervention,but the model group and normal group can not take any intervention measures,After modeling and after 14 days of intervention,the ethology of rats should be tested to observe the ethology effect of Mobi method on IBS visceral rats.3.3 pregnant rats were selected,and after their normal birth,the rats were randomly divided into a model group(n=20)and a normal group(n=10).After successful modeling,the model rats are randomly divided into experimental group(n = 10)and In the model group(n= 10)and the normal group of rats(n = 10),the experimental group adopted rubbing method intervention,but the model group and normal group can not take any intervention measures.After 14 days,colon pathological evaluation was performed to detect colon MC morphology and number.Transmission electron microscopy structure evaluation of colonic mucosa to clarify the effect of rubbing method on MC activation in IBS highly visceral rats.Results1.After 2 months of modeling intervention the rats in the normal group had white and bright fur,good mental state,more active activities,quick response,agility,and were not easy to be irritated.There was no thin stool or dry spherical stool.Rats in the model group had darker fur,less gloss,poorer mental state,less active behavior,prone to lying down,and were easily irritated.Stools were more sparse and unshaped.in the model group,It was found that the AWR scores of the rats in the model group under different pressures were higher than the scores in the normal group,the AWR score of the two groups of rats was statistically significant,P <0.05,and in the water avoidance experiment,the number of feces in the model group was more than that in the normal group.After statistical comparison,it was found that P<0.05 was statistically significant.indicating that the model group was modeled success.2.By examining the AWR scores of rats and the behavioral performance of the open field experiment,observing and counting the scores of the experimental group,model group and normal group,it was found that the AWR scores of the normal group and model group and the scores of the open field experiment did not change significantly,while the experiment The rats in the group had a significant increase in the open field test score after 14 days of intervention by the mobi method,and the AWR score decreased significantly,and the model group compared with the experimental group,P<0.05,and the model group compared with the normal group,P<0.05,with The statistical difference shows that the interventional method of rubbing abdomen can improve the visceral sensitivity state and behavioral behavior of IBS visceral hypersensitive rats.3.By observing the colon tissue of the rat,colon MC morphology and MC number and colon mucosal transmission electron microscopy evaluation,it was found that after the intervention of abdominal manipulation,The damage degree of the colon mucosa of the experimental group is better than that of the model group,but it is more serious than the normal group,The morphological structure of colonic mucosal epithelial cells in the experimental group was observed under transmission electron microscope more regularly and continuously than that in the model group.indicating that the intervention of rubbing method can help repair the colon pathology of IBS visceral highly sensitive rats damage.Compared with the model group,the number of MC in the colon decreased significantly in the experimental group,P <0.05,compared with the model group,the normal group had P <0.05 all had statistically significant differences indicating that the interventional methods of rubbing method can inhibit the activation and proliferation of MC in highly visceral IBS rats.Conclusions1.It is clear and feasible to adopt chronic mother-infant separation method combined with chronic mild stress method to model IBS visceral highly sensitive rats,and it can be applied repeatedly.2.Rubbing method can improve the visceral sensitivity state and behavioral performance of IBS visceral highly sensitive rats.3.Rubbing method can promote the repair of colon injury of IBS visceral highly sensitive rats,and can inhibit the activation and proliferation of intestinal mucosa MC.