| Objective:Establish the IBS-D rat models by acetic acid enema+bundle binding method;and to treat diarrhea-type irritable bowel syndrome rats with herb-partitioned moxibustion and investigate the effects of herb-partitioned moxibustion on expression of 5-HT ?5-HT3 R and 5-HT4 R proteins in rat colon tissue.Methods:Sixty male SD rats of SPF grade were divided into control group(12 rats)and model group(48 rats)after one week of adaptive feeding.The rats in the model group were subjected to acetic acid enema for 7 days and then restrained stress for 2weeks.Rats of the original module were randomly divided into model group,moxibustion group,Chinese medicine-medicine moxibustion group and western medicine group,with 12 rats in each group.In the normal group,no intervention was given.The model group was bundled and not treated.The Moxibustion group take a direct moxibustion.The herb-partitioned moxibustion group was treated with herb-partitioned moxibustion treatment,and the western medicine group was given a pimevir bromide tablet.After 2 weeks of treatment,the shape of Bristol stool was scored.The general behavioral changes of rats were observed,changes of fecal observation?trait score?water content,visceral sensitivity score.HE staining was used to observe the pathological changes of colon tissue.Elisa method was used to detect the expression of 5-HT in colon tissue,and the expression of 5-HT3 R and 5-HT4 R in colon tissue was detected by Western blot.Results:1.General behavioral changes in rats: The normal group of rats had a good mental state,a good living wave,a shiny coat and a rapid weight gain.Compared with the normal group,the model group was apathetic,with poor mobility,slow response,lack of luster in the fur,often curled up in the corner of the cage,and the weight gain wasslow.Compared with the model group,the spirit,activity,fur gloss and weight gain of the rats in the direct moxibustion group,the medicine-partition moxibustion group and the western medicine group were improved.2.Changes of fecal observation?trait score? water content: in the normal group,the anus was clean,the feces were formed,and the fecal traits were scored.In the model group,the feces were soft or not formed,and there was more fecal adhesion at the anus,and the fecal trait score was significantly increased;Rats in the medicine-separated moxibustion group and the western medicine group formed stools,which were softer and the fecal trait score was significantly lower(p<0.01).Compared with the normal group,the water content of the feces in the model group increased significantly(p<0.01),and the water content of the feces in the rats in the separated moxibustion group and the western medicine group decreased significantly compared with the model group(p<0.01).3.Visceral sensitivity score: Compared with the normal group,the measured model group rats had a lower AWR score of 3(the abdominal back muscles contracted strongly and lifted the abdomen off the ground),which was associated with increased visceral sensitivity in the model group.After the intervention of the medicine cake moxibustion and the pevimonium bromide,the water injection increased,indicating that the separate cake moxibustion and pivalvonium bromide have a good benign adjustment effect on the visceral hypersensitivity state of IBS-D rats.4.Pathological examination of Colon tissue: HE staining results of each group showed normal colon tissue structure and intact mucosal epithelium.Histopathological changes such as edema and ulceration,no inflammatory cell infiltration.5.Indicator detection: Compared with the normal group,the expression of 5-HT in the colon tissue of the model group increased(p<0.01),the expression of 5-HT3 R increased(p<0.05),and the expression of 5-HT4 R decreased(p<0.05).After treatment with direct acupuncture,separate cake moxibustion,and western medicine,the expression of 5-HT decreased(p<0.01),the expression of 5-HT3 R decreased(p<0.05),and the expression of 5-HT4 R increased.p<0.05).Conclusion:1.Rats were stimulated with acetic acid enema and bundled restraint,and the IBS-D rat model was successfully replicated after 3 weeks of intervention.2.The expressions of 5-HT,5-HT3 R and 5-HT4 R in the colon tissue of the model group were significantly different,indicating that 5-HT,5-HT3 R and 5-HT4 R were associated with the pathogenesis of IBS-D.3.The diarrhea symptoms and visceral hypersensitivity state of IBS-D rat model prepared by acetic acid enema combined with restraint stress stimulation have a benign regulation effect;and regulate the expression of 5-HT,5-HT3 R,5-HT4 R protein in rat colon tissue. |
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