| BackgroundIn recent years,the incidence and prevalence of diarrhea-predominant irritable bowel syndrome have been increasing,and it has become a global Gastrointestinal diseases.However,the etiology and pathogenesis of IBS have not been elucidated yet.In terms of treatment,western medicine still lacks effective treatment measures,mainly for the treatment of symptoms.In controlling the disease condition,preventing the recurrence of IBS-D,Chinese medicine has significant advantage,which adapts to the climate in treating IBS-D,especially its treatment based on syndrome differentiation.As Professor Li’s clinical experience prescriptions through his clinical practices,Tongxie Anchang decoction can significantly improve the syndromes of abdominal pain,loose stool,abdominal distension of IBS-D patients.It has good effect in treating IBS-D,but the specific mechanism is not so clear.ObjectiveIn this study,IBS-D rats were used as research objects to clarify the effect of Tongxie Anchang Decoction on them.Taking PLC-γ/TRPV1 pathway as the starting point,to clarify whether Tongxie Anchang Decoction treated IBS-D by regulating the PLC-y/TRPV1 pathway,to clarify the target of Tongxie Anchang Decoction in treating IBS-D,and then to provide experimental evidence for further clinical application of Tongxie Anchang Decoction in treating IBS-D.MethodsExperiment one:The IBS-D rat model was established by given enema with glacial acetic acid,colorectal dilation,tail-clamping stimulation and senna gavage.After modeling,the rats’ general situation,diarrhea situation,Abdominal withdrawal reflex score were measured.After treating,the above indicators were tested again.Experiment two:The IBS-D rat model was established by given enema with glacial acetic acid,colorectal dilation,tail-clamping stimulation and senna gavage.After 2-week intervention,PLC-y/TRPV1 gene and protein expression were detected by Real time-PCR and Western-blot,Elisa was used to detect the expression of SP and CGRP in serum.ResultsExperiment one:1 From the third day of model building,in addition to the normal group,with the change of time,the mental state of the rats became sluggish with sensitivity,the appetite was poorer,the hair was messy and gradually lost its luster.By the 4th week,the rats had very poor mental status,with dry and falling hair,or even a complete loss of luster,weight loss,and activity is extremely poor.After intervention of Tongxie Anchang Decoction,compared with the model group,the rats in each treatment group had gradually improved their spirits and began to gain weight.The appetite recovered,the hair color began to become shiny,and the activity was good.2 After the start of modeling,rat stool became soft and gradually pulpy;after 4-week modeling,the fecal Bristol score of rats was higher than the normal group(P<0.01),the fecal water content increased significantly(P<0.01);after the intervention of Tongxie Anchang Decoction,the rats’ fecal Bristol score decreased(P<0.01).3 After establishing the model,the AWR score of rats was higher than that of the normal group(P<0.05,P<0.01).After the treatment of Tongxie Anchang Decoction,the AWR score of rats decreased(P<0.05,P<0.01).Experiment two:1 The results of Real time-PCR showed that the expression of PLC-y mRNA and TRPV1 mRNA in colonic tissue of rats with IBS-D was increased(P<0.05),the expression of TRPV1 mRNA was decreased in different dose groups(P<0.05),the expression of TRPV1 protein was increased in model group(P<0.05),and the expression of TRPV1 protein in colon decreased significantly(P<0.01,P<0.05)after the intervention of Tongxie Anchang Decoction.2 In the model group,the contents of SP and CGRP in serum increased significantly(P<0.01),and the contents of SP and CGRP in the serum of rats treated with Tongxie Anchang Decoction decreased significantly(P<0.05,P<0.01).Conclusion1 After the establishment of the model,the intestinal tract of rats was highly sensitive,the fecal Bristol grade and fecal water content were increased,and the symptoms of abdominal pain and diarrhea were in line with the clinical characteristics of IBS-D.IBS model could be successfully duplicated by tail stimulation combined with Senna leaf.2 Tongxie Anchang Decoction can reduce the AWR score,the stool Bristol grade and the fecal water content of the IBS-D model rats,which indicates that the Tongxie Anchang Decoction can reduce the visceral hypersensitivity of IBS-D rats and improve the diarrhea symptoms of the rats.3 Tongxie Anchang Decoction decreased the expression of PLC-γ、TRPV1 genes and proteins in colon tissue,and decreased the volume of SP and CGRP in serum,which suggested that Tongxie Anchang Decoction could inhibit visceral hypersensitivity of IBS-D rats by down-regulating the function of PLC-γ/TRPV1 signaling pathway. |
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