| Purpose:1、Studying modeling method of animal of the model of Irritable bowel syndrome (IBS)visceral hypersensitivity. And the stability of the model after successful model is observed, which provides the basis for further experiments.2、establishing the model of Irritable bowel syndrome (IBS)visceral hypersensitivity, and to detect the expression of genes in the hippocampus and the intestinal mucosa of rats in each group, To explore the possible mechanism of IBS-D mediated pathway of visceral hypersensitivity,Also to provide theoretical basis for the clinical development of drugs and for the study of the mechanism of traditional Chinese Medicine3、according to the differential gene expression, we study the mechanism of Tongxieyaofang relief visceral hypersensitivity in IBS-Dmethod:1、Distening the colorectal in young rats and adult rats With different sizes of angioplasty balloon. Detecting the visceral sensitivity and pain threshold in rats after 2 weeks of cessation of stimulation, According to the results of the blank group comparison to determine whether the success of modeling. The pain threshold were detected again after 4 weeks and 6 weeks after stimulation. According to the results of the comparison of the two pain threshold values, the stability of the two modeling methods was evaluated, to determine whether this modeling method is consistent with the characteristics of irritable bowel syndrome.2、The SD rats of eighth days were selected as the experimental subjects, Rats in the experimental group received angioplasty balloon dilation (pressure of 60mmHg) 8-21 days after birth, Stimulate 1 times a day, each lasting for 1 minutes. Control group of rats were gently grabbed, touch the rat will be 1 minutes. Growth to 25 days later, rats and mice will be separated. Rats in the completion of the above stimulus, no longer give any stimulation. For fifth weeks, the visceral sensitivity of rats was evaluated. According to the results of the two groups, the success of the visceral hypersensitivity model was judged. The rats in the group of tongxieyaofang were poured into the stomach of the traditional Chinese medicine(tongxieyaofang) for four weeks. Detecting the visceral sensitivity of rats again, and judging whether the traditional Chinese medicine is effective according to the results. After stopping the drug to continue the normal feeding for 1 weeks. Then take the rat serum, hippocampus, intestinal mucosa samples, the samples were sent to Shanghai Biotechnology Corporation Gene expression in brain of model group and blank control group were analyzed by gene chip technology, Immunohistochemistry technique was used to detect the expression of protein.Result:1、Compared with the adult rat model, the animal model established by the Neonatal rats is more stable. To a certain extent, the animal model established by neonatal rats as the experimental object is more in line with the characteristics of irritable bowel syndrome.2、Gene differential expression, The expression of c-fos gene in hippocampus of model group was significantly higher than that in control group (p<0.05). The expression of c-fos gene in the hippocampus of the rats in the group of TongXieYaoFang was significantly lower than that in the model group in the hippocampus of the model group (p<0.05).3、Gene differential expression, The expression of ILlrl in the model group was significantly higher than that in the control group (IL1r1) in the control group (p<0.05),The expression of ILlrl gene in the model group was significantly lower than that in the p<0.05 group (IL1r1).4、Gene differential expression, Expression of Tmeff2 gene in model group was significantly higher than that in the intestinal expression in the control group (P>0.05).5、Gene differential expression, There were no significant differences between the two groups in the expression of BDNF gene in the rat intestinal mucosa tissue samples, and the expression of TRKB receptor gene was not significantly different between the groups. There was no significant difference (p>0.05) between the two groups in the expression of BDNF gene in the rat hippocampal tissue samples, and the expression of TRKB receptor gene was not significantly different between the groups. |
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