| Objective: To establish a IBS-D animal model and study the changes of AQP3,Claudin-1,mast cell and TNF-a concentration in the colon of IBS-D model rats,and to explore the mechanism of the intervention of IBS-D by different doses Tong –Xie-Yao –Fang(TXYF).Methods: Forty-eight adult SD rats were randomly divided into normal group(the four are female,the four are male,which are not to participate in modeling)and model group(the rest are fourty),that are made by Chronic restraint stress combined with constaint method.After successfuy moding,the model group were grouped into model control group,pinaverium bromide group,TXYF(low,middle,high)tradition Chinese medicine group.rats in each group were given intragastric administration of different drugs or 0.9% saline for 7 days,once a day.After the treatment,The proximal colon tissues were to be stained with HE and counted mast cells,The expression of AQP3 and Claudin-1 and the detection of TNF-a concentration were observed in distal tissues of rats.Results: 1.Model evaluation: in addition to the normal group,the model group rats had low coat color,even vertical hair,decreased activity,decreased appetite,likes to curl up,pile up,slow reaction,soft or irregular stool.The body weight increased slowly(P < 0.05).intestinal pain area decreased(P < 0.05),stool Bristol score(P < 0.05).Pathological sections showed that the colonic mucosa was intact without interstitial hyperemia edema and occasionally inflammatory cell infiltration in each group which indicated that the model was successful.2.The effect of TXYF on MC in proximal colon tissue of IBS-D model rats: compared with the normal group,the number of mast cells in the proximal colon tissue of the model control group increased(P < 0.05),and compared with the model control group,the number of mast cells in the proximal colon tissue of the model control group increased significantly(P < 0.05).The number of mast cells in the proximal colon of rats in the piveronium bromide group(low,medium,high)dosage group decreased significantly(P < 0.05).3.The effect of TXYF on TNF-a in distal colon tissue of IBS-D model rats: compared with the normal group,the content of TNF-a in the model control group was significantly higher than that in the mormal group,The difference was statistically significant(P < 0.05).compared with the model control group,the content of TNF-a in distal colon tissue of rats in the group of piveronium bromide and the group of TXYF(middle and high dose)were all decreased.The difference was statistically significant(P < 0.05).4.Effect of TXYF on expression of Claudin-1 mRNA and AQP3 mRNA in distal colon of IBS-D rats: compared with normal group.The expression of AQP3 mRNA and Claudin-1 mRNA in the distal colon of the model group was significantly lower than that in the normal group(P < 0.05).Compared with the model control group,the TXYF(middle,high)dose group and piveronium bromide group could improve the diarrhea state of the model rats and up-regulate the AQP3 mRNA and Claudin-1mRNA of the proximal colon of the rats,That was significantly different(P < 0.05).Conclusions: 1.The IBS-D rats model was established successfully.2.IBS-D rat colon MC,AQP3,Claudin-1 and the level of TNF-a are abnormal expression,which suggests that colon tissue MC,AQP3,Claudin-1 and the TNF-a level be involved in the prescription.3.TXYF can reduce the number of MC in the pathogenesis of IBS-D and up regulate AQP3 mRNA,Claudin-1mRNA expression level and decrea levels of TNF-a,to promote colon on intestinal cavity water reabsorption and repair of the intestinal mucosal barrier,to treat diarrhea.4.Regulating AQP3/Claudin-1 may play a role on the mechanism that TXYF can treat IBS-D. |
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