| Objective:By constructing the rat IBS-D model,the changes of 5-HT in serum,colon tissue and brain tissue and the expression of SERT in colon were studied.To explore the mechanism of Tongxie Anpi Decoction in the treatment of diarrhea-predominant irritable bowel syndrome,and to provide experimental evidence for the treatment of IBS-D in Tongxue Anpi Decoction.Methods:After 48 weeks of adaptive feeding of 48 SD rats,48 male and female SPF SD rats were randomly divided into 12 groups and 36 model rats(model group,Chinese medicine).Group,western medicine group,12 in each group,the animal model of IBS-D liver spleen deficiency was established by restraining + senna leaf gavage method.In the first to fourth weeks of modeling,the senna leaf decoction was administered to the stomach.From the third week,the rats were placed in a plastic bottle of about 550 ml in size,so that the rats could only move slightly in the bottle before and after the movement for 3 hours./ time / day,the start time of each bond is not fixed.In the four weeks of modeling,each rat was intermittently trapped for 1 minute,so that it was good to make a mourning.After 4 weeks of modeling,the rats in each group were evaluated for general condition,rat body weight,stool traits,visceral sensitivity and liver stagnation,and 2 rats were randomly selected from each group for pathology.The test determines that the modeling is successful.After successful modeling,the normal group and the model group were given normal saline,the western medicine group was given Alosetron and the Chinese medicine group was given Tongxie Anpi Decoction for 2 weeks,the animals were treated,the experimental specimens were selected,and the ELISA method was selected.The serum 5-HT content was detected.The levels of 5-HT and colonic SERT in colon tissue and brain tissue were detected by immunohistochemistry.The expression of 5-HT protein in colon tissue was detected by Western blot.Results:1.General situation: Before the model was made,the hair of the rats was shiny,full of spirits,stalking around,and the stool was granular.At the beginning of the modeling,when the rat restraint frame was restrained,it resisted strong and biting.After the model was established,the rats in the model group were lazy,the diet was reduced,the degree of diarrhea increased,and the response to external stimuli was slow.The degree of resistance and biting during restraint was weaker than before the modeling;the body gained slowly and the hair was dirty and messy.Lack of shine.The normal group had the same mental state,diet,stool,and color before the modeling,and the body shape was rounder before the modeling.2.Changes in body weight of rats: After modeling: Compared with the normal group,the growth rate of each module was significantly lower(P<0.05),and the difference between the modules was smaller(P>0.05).The body weight growth rate of the model group was significantly higher than that of the model group(P<0.05).There was no difference between the alosetron group and the traditional Chinese medicine group(P>0.05).3.Changes in rat feces: After modeling: the fecal trait scores of the rats in each group were significantly higher than those in the normal group(P<0.05);the differences between the groups were small(P>0.05).Compared with the model group,the rate of diarrhea in rats was significantly lower than that in the model group(P<0.05).There was no difference between the alosetron group and the traditional Chinese medicine group.The effect of loszecon in the treatment of diarrhea in IBS-D rats was obvious,and the therapeutic effect was similar.4.The changes of the retraction and reflex of rat abdominal wall: after modeling: the AWR scores of the rats in each group were significantly higher than those in the normal group(P<0.05);there was no difference between the groups.The visceral sensitivity of rats treated with Alosetron group and Tongxie Anpi Decoction group was significantly lower than that of the model group(P<0.05).5.The changes of the scores in the experimental field of rats: after modeling:the scores of open field experiment in each model group were significantly higher than those in the normal group(P<0.05);the differences between the models were small(P> 0.05).After treatment: compared with the model group,the scores of open field test in the azosetron group and the Chinese medicine group were significantly increased(P<0.05),and the alosetron group was compared with the traditional Chinese medicine group(P<0.05).The difference was statistically significant.significance.Both Tongxie Anpi Decoction and Alosetron can reduce the liver stagnation status of IBS-D rats,but the effect of Tongxie Anpi Decoction is better than that of alosetron.6.Sample 5-HT test results6.1 Serum ELISA results: Compared with the serum 5-HT content in the normal group,the serum 5-HT content in the model group increased significantly(P<0.05).Compared with the serum 5-HT content of the model group,the Tongxie Anpi Decoction group and the alosetron group were significantly decreased,with significant statistical difference(P<0.01).Compared with the western medicine control group,the difference between the traditional Chinese medicine treatment group and the western medicine control group was small(P>0.05),and there was no statistically significant difference.6.2 Western blot analysis of colon tissue: Compared with the model group,the expression of 5-HT protein in the colon of Tongxie Anpi Decoction group and Alosetron group was significantly lower(P<0.01).The normal group was compared with the model group.The 5-HT content was significantly increased(P<0.01).The difference between the Chinese medicine treatment group and the western medicine control group was small(P>0.05),and there was no statistically significant difference.6.3 Immunohistochemical results of colon and brain tissue(5-HT):Compared with the normal group,the level of 5-HT in the model group increased significantly(P<0.01);after the treatment of Tongxie Anpi Decoction and Alosetron The level of 5-HT was significantly lower than that of the model group(P<0.01).There was no significant difference between Tongxie Anpi Decoction and Alosetron group(P>0.05).6.4 Colon Immunity(SERT)Results:SERT was low in the intestinal mucosa of the model group,and was significantly lower than the normal group(P<0.01).The expression of SERT in the intestinal tissue of the model group was significantly higher than that of the model group(P<0.01).Intestinal tissue SERT was also significantly higher after treatment(P<0.01).There was no significant difference between the Chinese medicine group and the western medicine group(P>0.05),which was not statistically significant.Conclusion:1.Tongxie Anpi Decoction can significantly improve the liver stagnation and spleen deficiency in rats,promote the weight gain of rats,improve the symptoms of diarrhea in rats,and reduce the intestinal sensitivity of rats;2.The pathogenesis of IBS-D may be associated with increased levels of5-HT,decreased levels of SERT,resulting in decreased visceral pain threshold and increased intestinal sensitivity;3.The treatment of IBS-D by Tongxie Anpi Decoction may regulate the levels of 5-HT and SERT,attenuate the excitability of spinal cord neurons,thereby increasing the visceral pain threshold and reducing intestinal hypersensitivity. |
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