Mechanism Research Of Tongbian Decoction In The Treat Of Slow Transit Constipation Based On The Protein Kinase A(PKA) Conduction Pathway

Objective:In this study,We used the compound diphenoxylate to establish the slow transit constipation model,followed the pathegeny and pathogenesis theory of"Deficiency of body fluid,the boat stops",observed changes of neurotransmitters in the model’s intestinal tissues and analyzing the pathological process of intestinal dryness,in order to elucidate the biological basis of related disease.We used the famous prescription "Tongbian Decoction”to intervene the animal model.At final,from the perspective of modern medicine,to explain the science of Chinese medicine treatment which named nourish yin of intestinal disease.Methods:60 rats were randomly divided into the normal group(20 rats)and the model group(40 rats),and the slow transfer constipation model was replicated by compound diphenoxylate.Randomly selected normal group and model group with 10 rats in each observation to observe the intestinal transmission function,and used immunohistochemistry,Western-blot and fluorescence quantitative PCR for detection of colonic mucosal PKA signaling pathway,AQP3,AQP4 distribution,expression and relative content.Results:1.After the modeling,rats of model group compared with normal group,the amount of feces was lower,the volume was smaller,the quality was harder,the water content was significantly less,the propelling rate of carbon and ink was significantly reduced,the number of residual feces in the colon was significantly higher.2.When the model was established successfully.The results of immunohistochemistry showed that compared with normal group,the expressiono of AQP3 of model group was significantly increased(p=0.000);compared with normal group,the expressiono of AQP4 of model group was increased(p=0.028);compared with normal group,the expressiono of PKA of model group was significantly increased(p=0.000).3.After 4 weeks of drug intervention,the general condition of the rats in the"Tongbian Decoction" group and "Tongbian Decoction"+H89 group was improved.,there was no significant difference between two groups.The number and the water content of the feces increased significantly(p<0.001).The amount and water content of feces of the other three groups were significantly higher than those in the group(p<0.001).Compared with the normal group,there was no significant difference in the amount of feces in the "Tongbian Decoction" group and the "Tongbian Decoction"+H89 group,the water content was more than that of the normal group.4.After 4 weeks of drug intervention,immunohistochemistry,WB and Q-PCR results suggested that the expression of AQP3,AQP4 and PKA were in the model group was the highest,followed by "Tongbian Decoction" group,"Tongbian Decoction"+H89+H89 group,normal group.Conclusion:1.The use of compound diphenoxylate could successfully replicate the stable slow transit constipation rat model with high safety and conformed to the clinical features of the disease.The high expression of AQP3,AQP4 and PKA signaling pathways may be the biological basis for the pathogenesis of STC.“Tong Bian" decoction can significantly improve the general situation of STC model rats,increase the number,quality and water content of feces,promote intestinal peristalsis and reduce the residual feces of colon."Tong Bian" decoction can effectively treat STC."Tong Bian" decoction is by blocking the expression of PKA signaling pathway,thus reducing the expression of AQP3 and AQP4 and reducing the absorption of water to achieve the effect of the treatment of STC.