Expression Of ICC And SCF/c-Kit Pathway In Rat With Slow Transit Constipation Treated By Yi Qi Run Chang Recipe

Slow transit constipation is a common type of functional constipation,accounting for16%-40%of chronic constipation. Its cause is unknown.lt has no obvious pathological changes, difficult treatment, recurrent symptom, which seriously affects the quality of life of patients and is easy to induce other diseases. Research and treatment of this disease has important significance. Based on efficacy evaluation showed that Yi Qi Run Chang Recipe is a safe and effective treatment of functional constipation,now our subject is to seeks to further explain the mechanism of Yi Qi Run Chang Recipe treatment of STC by animal experiment.Objective:To study colon ICC changes and SCF/c-Kit pathway affect with STC after treatment with Yi Qi Run Chang Recipe to reveal the mechanism.Method:48Wistar rats were randomly divided into normal control group, model control group,Qi Rong Run Chang oral Liquid group and Yi Qi Run Chang Recipe group. Normal control group were not modeling, the rest would be orally administered compound diphenoxylate for120days, so STC can be induced in rats. After the modeling, the control group received normal feeding.The Model control group, Qi Rong Run Chang oral Liquid group and Yi Qi Run Chang Recipe group were fed with an equal volume of saline, Qi Rong Run Chang oral Liquid dilution and Yi Qi Run Chang Recipe Liquid respectively with20days of continuous perfusion. After the last dosing, observe the general situation and defecation,count the number of fecal pellets in24hours with metabolic cages.Then the rats were fasted24hours,receive activated carbon suspension by intragastric administration.Kill them30min later by cervical dislocation. Calculated the percentage of carbon propellin [(Distance from the tip of powder to the pylorus/full length of gut) x100%]in relaxed state. Cut two colonic circular muscle tissues about1cm then fixed in neutral formalin solution and stored in EP tube for each rat.After the use of HE staining,observe the rat colon tissue.After the use of immunohistochemical staining,analyze different parts of the ICC’s area and integrated optical density with the application of Image-Pro Plus6.0software. The c-kit and SCF gene expression was observed by RT-PCR and calculated by means of the2-△△Ct method.Result:1.The defecation grains in24hours:Compared with the model control group, it significantly increased in Yi Qi Run Chang Recipe group and increased in Qi Rong Run Chang oral Liquid group. Compared with the normal control group, the defecation grains in24hours in Yi Qi Run Chang Recipe group had increased, but not statistically significant by statistical analysis.2.Carbon powder advance rate:Yi Qi Run Chang Recipe group compared with the model control group, it improved significantly. Compared with Qi Rong Run Chang oral Liquid group, the Carbon powder advance rate in Yi Qi Run Chang Recipe group had increased, but not statistically significant by statistical analysis,as well as to normal control group.3.Colonic ICC area and IOD:There was a statistically significant difference between normal control group and the model control group, the former was significantly higher in ICC area and IOD.For the Yi Qi Run Chang Recipe group and the model control group, there was a statistically significant difference between them, the former had an increase in the area and IOD of ICC.There was no statistically significant difference between Yi Qi Run Chang Recipe group and normal control group,but the former’s area and IOD was also higher.For Qi Rong Run Chang oral Liquid group and the model control group, there was a statistically significant difference, the former’s ICC area and IOD was significantly higher than the latter. Yi Qi Run Chang Recipe group,compared with Qi Rong Run Chang oral Liquid group,had a significant difference. The former ICC area and IOD was higher than the latter.4.SCF mRNA and c-kit mRNA:Compared with the normal control group, the model control group’s SCF mRNA expression levels relatively reduced.SCF mRNA expression of Yi Qi Run Chang Recipe group and Qi Rong Run Chang oral Liquid group was relatively elevated compared with the model control group. Although without statistically significant, Yi Qi Run Chang Recipe and Qi Rong Run Chang oral Liquid might make SCF mRNA expression levels close to normal levels.Compared with the normal control group, model control group’s c-kit mRNA expression level was relatively lower. Compared with the model control group, c-kit mRNA expression of Qi Rong Run Chang oral Liquid group and Yi Qi Run Chang Recipe group was relatively elevated. The latter’s degree representing the improving of c-kit mRNA expression levels was more than the normal control group.Conclusion: Yi Qi Run Chang treatment for STC rats can promote intestinal emptying. The possible mechanism is increasing the expression of SCF mRNA and c-kit mRNA,affecting the SCF/c-Kit signaling pathway, raising the number of the colon ICC, thus promoting intestinal motility.