| Objective:To investigate the mechanism and dose-effect relationship of Tongbian Decoction in treating STC by observing the autophagy of ICC in STC rat model.Methods:80 SD rats were randomly divided into blank group(20 rats)and model group(60 rats).STC rat model was replicated with compound phenylethylpiperidine.Ten rats in the blank group and 10 rats in the model group were randomly selected.The general condition(body weight,food intake,water intake),24-hour total feces and water content,intestinal transmission function,colon myoelectricity were observed.The expression of ICC and c-kit was detected by immunohistochemistry,protein immunoblotting and PCR.After 1 week of drug withdrawal,the total amount of feces and water content of rats in the model group were measured 24 hours after 1 day and 7 days after the end of modeling.After successful replication of STC rat model,the remaining 50 rats in STC rat model group were randomly divided into model group 10 rats,Tongbian Decoction low,medium and high dose group 10 rats,Mosapride group 10 rats.After 6 weeks of drug intervention,the general situation of rats(body weight,food intake,water consumption),24 h fecal total and water content,intestinal transmission function,colon myoelectricity were observed,and ICC was observed by transmission electron microscopy.The expression of c-kit,Beclinl and LC3 were detected by Western blotting and PCR.Result:1.Compared with the blank group,the general condition of rats in the model group was worse,the total feces volume and water content in 24 hours decreased(P<0.01),the rate of carbon powder propulsion decreased(P<0.01),the number of colon retained feces increased(P<0.01),the variation of EMG frequency and amplitude increased(P<0.01),and the expression of ICC and c-kit decreased(P<0.01).After 1 week of withdrawal,the total amount of feces and water content of rats in the model group did not change significantly(P>0.05).2.Compared with the blank group,the structure of ICC cells in the model group changed significantly and autophagy increased.Compared with the model group,after 6 weeks of Tongbian Decoction treatment,the body weight,food intake,water consumption,total feces and water content increased(P<0.01),the propelling rate of charcoal powder increased(P<0.01),and the retention of feces in colon decreased(P<0.01).The frequency and amplitude variation of EMG decreased(P<0.01),the expression of ICC and c-kit increased(P<0.01),the expression of Beclinl and LC3 decreased(P<0.01),and there was significant difference among low,medium and high dose Tongbian Decoction groups.The effect of high dose Tongbian Decoction was significant.3.Compared with the model group,the rats in Mosapride group had increased body mass,increased food intake,decreased water consumption(P<0.01),increased total feces and water content(P<0.01),increased carbon powder propelling rate(P<0.01),decreased colon retained feces(P<0.01),decreased electromyogram frequency and amplitude variation(P<0.01),but no significant changes were observed in ICC and c-kit expression(P>0.05),and Bec-kit expression(P>0.01).The expression of linl and LC3 did not change significantly(P>0.05).Conclusion:1.The STC rat model was successfully reproduced with compound phenylethylpiperidine,and the STC rat model was stable and safe.2.Tongbian Decoction can significantly improve STC symptoms.The main mechanism may be related to down-regulation of Beclinl and LC3 expression,inhibition of excessive autophagy of ICC,up-regulation of c-kit expression,promotion of ICC repair,stimulation of intestinal peristalsis and increase of water content.3.Tongbian Decoction has a certain relationship with the dosage of STC,and high-dose Tongbian Decoction has the best effect on STC.4.Mosapride can promote intestinal peristalsis,increase the total amount of feces and water content,and increase intestinal transmission function to treat STC by improving the general condition of rats(body weight,food intake and water intake),but it does not inhibit the excessive autophagy of ICC. |
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