| Objectives:To explore the possible mechanism of treating STC with Bensatong Mixture by observing the effect of Biansetong Mixture(BST)on SCF/c-Kit signaling pathway and colonic myoelectricity activity in STC model rats induced by loperamide,so as to provide a more comprehensive experimental basis for the treatment of STC with Bensatong Mixture.Methods:40 healthy male SD(Sprague Dawley)rats,were divided into two groups according to the principle of random grouping:10 in normal control(NC)group and 30 in model group.The STC rat model was replicated with loperamide.After 30 days of modeling,the grain number of stool,the water content of stool at 24h and first black stool time were compared between the NC group and the modeling group to verify whether the model replication was successful.After successful modeling,30 rats in the model group were randomly divided into three groups:Model Control group(MC),mosapride group(Positive Control(PC)),and Biansetong Mixture group(BST).After 14 days of drug intervention,the general situation of rats in each group were observed,the grain number of stool was counted,the water content of stool for 24 hours and the small intestine promoting rate of carbon ink were measured,the myoelectricity activity of colonic smooth muscle of rats was detected by the biological function experiment system,and the expression of SCF and c-Kit was detected by Immunohistochemistry and RT-PCR.Results:1.After 30 days of modeling,compared with the NC group,the grain number of stool and the water content of stool decreased at 24h,and the first black stool time was prolonged(P<0.01).2.After 14 days of drug intervention,compared with the NC group,the grain number of stool and the water content of stool at 24h,small intestine prompting rate of carbon ink of rats in the MC group declined(P<0.01).Compared with the MC group,the grain number of stool and the water content of stool at 24h,small intestine prompting rate of carbon ink in the BST group and PC group increased(P<0.05).The BST group was better than the PC group in improving the water content of stool at 24h(P<0.05),but there was no significant difference in improving the grain number of stool at 24h and small intestine prompting rate of carbon ink(P>0.05).3.The results of Immunohistochemistry showed that protein expressions of SCF and c-Kit in the colon tissues of rats in the MC group were decreased compared with the NC group(P<0.05).Compared with the MC group,protein expression of SCF and c-Kit increased in the colon tissues of rats in the BST group(P<0.05).Compared with the MC group,SCF and c-Kit protein expression in the colon tissues of rats in the PC group showed no significant changes(P>0.05).RT-PCR results indicated that the mRNA expression levels of SCF and c-Kit in the colon tissues of rats in the MC group were significantly lower than those in the NC group(P<0.05).Compared with the MC group,the mRNA expression levels of SCF and c-Kit in the colon tissues of rats in the MC group were significantly increased after BST intervention(P<0.05),while the mRNA expression levels of SCF and c-Kit in the colon tissues of rats in the PC group were not significantly changed(P>0.05).4.Colonic electromyography(EMG)showed that compared with the NC group,the slow wave frequency of the colon slowed down,the amplitude increased,and the variation coefficient of frequency and amplitude increased in the MC group,with statistically significant differences(P<0.05).In the BST group and the PC group the frequency accelerated and the amplitude decreased,and the variation coefficient of frequency and amplitude declined compared with the MC group(P<0.05).No significant difference was found in the frequency,amplitude and the variation coefficient of frequency and amplitude between the BST group and the PC group(P>0.05).Conclusion:1.The STC rat model is replicated by loperamide,which has a certain time stability and is suitable for STC experimental study.2.BST could increase the grain number of stool,the water content of stool and intestine prompting rate in tin STC model rats,thus improving constipation symptoms.3.The mRNA level and protein expression of SCF and c-kit in the colon tissues of STC model rats decrease,and the mRNA level and protein expression of SCF and c-Kit in the colon tissues of STC model rats increase synchronously after BST intervention.Therefore,we speculate that the BST in the treatment STC is related to the recovery of SCF/c-Kit signaling pathway.4.On the STC model,colon slow wave frequency decreases,amplitude,frequency and the amplitude variation coefficient increase,after the intervention of BST,the rat colon slow wave frequency increases,the amplitude decreases,and variation coefficient between slow wave frequency and the amplitude reduces,which suggest that BST can restore colon slow wave rhythmic and stability,and reduce the slow wave amplitude,and strengthen the function of intestinal contraction,promote the excretion of intestinal contents. |
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