| Objective:Based on previous studies,this paper further evaluated the clinical efficacy of Xiaoyao Decoction in treating liver depression and spleen deficiency IBS-D from both clinical and animal experiments.And to observe the effects of Xiaoxiao Decoction on vasoactive intestinal peptide(VIP),calcitonin gene-related peptide(CGRP),skeleton protein(Claudin-4),ZO-1 and Occludin in the colon tissue of IBS-D rats,so as to explore the mechanism of Xiaoxiao decoction in the treatment of IBS-D deep layer.To provide a new modern medical theory basis for the treatment of IBS-D with traditional Chinese medicine.Methods:1 Clinical study:Sixty patients who met the inclusion criteria were randomly divided into experimental group(30 cases)and control group(30 cases)according to the random method.The treatment course was 4 weeks.During the treatment,the control group was given Piviteronium bromide orally,and the experimental group was given Xiaoyao decoction orally on the basis of the control group.The score changes of TCM syndrome,IBS-SSS,SAS and SDS scales,as well as the differences of TCM clinical efficacy,recurrence rate and safety indexes between the two groups were compared before treatment and after 3 months of follow-up.2 Experimental research:In this study,48 healthy SD rats were randomly divided into blank group(n=12)and modeling group(n=36).The blank group was fed normally,and the modeling group was established to establish IBS-D rat model of liver depression and spleen deficiency by "senna leaf gavage + chronic restraint combined with tail clip stimulation",and the cycle was 2 weeks,and the evaluation was made after the cycle.After successful evaluation,the rats were randomly divided into model group,Xiaoyao decoction group and Piviteronium bromide group,with 12 rats in each group.The changes of various indexes were observed and recorded,and then the rats were given the corresponding drugs intragastric administration.After 14 days of continuous administration,the rats were killed.Real-time fluorescence quantization(RT-q PCR)was used to detect mrna expression of VIP,CGRP,ZO-1,Claudin-4 and Occludin in colon tissue of rats in each group.The content of VIP,CGRP,ZO-1,Claudin-4 and Occludin in colon tissue of rats were detected by immunohistochemical SP method.Results:1 Clinical part:(1)Both groups could reduce TCM syndrome score,and the experimental group had better efficacy(P<0.01);(2)IBS symptom severity scores in both groups were significantly decreased compared with before(P<0.01),and the downward trend in experimental group was more obvious(P<0.05);(3)The scores of SAS and SDS in both groups were decreased compared with those before treatment(P<0.01,P<0.05),and the anxiety and depression state in the experimental group could be effectively alleviated compared with the control group(P<0.05).(4)After the course of treatment,the effective rate of TCM syndrome in the experimental group was 93.3%,and the total effective rate in the control group was80.00%.(5)After 3 months of follow-up,the recurrence rate of the experimental group was10.00% and that of the control group was 26.67%.The long-term efficacy of the two groups was better than that of the experimental group(P<0.05).(6)During treatment,there were no adverse changes in the safety indexes of patients in both groups,which proved that the clinical application was safe and feasible.2 Experimental part:(1)General conditions of the rats: before modeling,the general conditions of the rats were normal;after modeling,compared with the blank group,the rats in the modeling group showed decreased activity,mental burnout,emotional irritability,dry and dark fur,and emaciation;Compared with the model group,after treatment,the spirit and activity of rats in each group were significantly improved,stool shape and diarrhea were significantly improved.(2)Changes in food intake: After the modeling,the food intake of rats in the modeling group was decreased compared with that before(P<0.05),and after treatment,the food intake of rats was significantly increased(P<0.05),and there was no difference between the groups after treatment(P>0.05).(3)Changes in fecal traits: After modeling,fecal Bristol score in the modeling group was significantly higher than that in the blank group(P<0.05).After treatment,fecal Bristol score of rats in all groups was decreased to varying degrees(P<0.05),and Xiaoyao Decoction was better than Xiaoyao decoction between the two groups(P<0.05).(4)Abdominal wall retraction reflex score(AWR): After modeling,the pain threshold of rats in modeling group was significantly lower than that in blank group(P<0.01);After treatment,compared with model group,pain threshold of rats in Xiaoyao Decoction group was increased(P<0.05),but no difference was found in Pivectium bromide group(P>0.05),and the effect of Xiaoyao decoction was better between the two groups(P<0.05),suggesting that this was related to the change of visceral sensitivity of model rats.(5)Sugar water preference experiment: After the modeling,the score of sugar water preference experiment of rats decreased compared with that of blank group(P<0.05),suggesting that the rats were in a state of anxiety and depression.(6)Colonic histopathological examination: colonic tissue sections of rats in each group indicated that the colonic mucosa was intact,without congestion or edema,with a small amount of local inflammatory cell infiltration,and no damage to the morphological structure.(7)RT-q PCR test results:Compared with model group,after treatment,ZO-1,Claudin-4and Occludinm RNA expression in colon of rats in 2 groups were up-regulated(P<0.0001),while VIP and CGRPm RNA expression in Xiaoxiaojian group were down-regulated(P<0.0001).The expressions of VIP and CGRPm RNA in Pivillium bromide group were down-regulated(P<0.01,P<0.0001);Compared with Pivillium bromide group,mrna expression of ZO-1,Claudin-4 and Occludin in Xiaoxiao decoction group was significantly up-regulated(P<0.01,P<0.001),mrna expression of VIP and Cgrpwas significantly down-regulated(P<0.0001).(8)Immunohistochemical SP test results: After treatment,compared with model group,positive expression of VIP and CGRP proteins in colon of rats in Xiaoxiaojiao decoction group was down-regulated(P<0.0001),positive expression of ZO-1,Claudin-4,Occludin protein was up-regulated(P<0.0001,P<0.0001,P<0.001).The positive expressions of VIP and CGRP proteins in colons of Pivillium bromide group were down-regulated(P<0.05,P<0.001),while positive expressions of ZO-1,Claudin-4and Occludin were up-regulated(P<0.001,P<0.0001,P<0.05).Compared with Pivillium bromide group,mrna expressions of VIP and Cgrpwere down-regulated in Xiaoxiaojian group(P<0.0001,P<0.05),and mrna expressions of ZO-1,Claudin-4 and Occludin were up-regulated in Xiaoxiaojian group(P<0.001,P<0.05,P<0.05).Conclusions:1.This clinical study confirmed that the clinical effect of Xiaoyao Decoction combined with Pivillium bromide in the treatment group of patients with liver depression and spleen deficiency IBS-D and the improvement degree of anxiety and depression state were significantly better than that of the single treatment group of Pivillium bromide,and the long-term effect of the trial group was better than that of the control group.2.The modeling method of "senna leaf gavage + chronic binding stress combined with tail clip stimulation" was adopted in this experiment,and the results were in line with the standard animal model of liver depression and spleen deficiency IBS-D,indicating successful modeling.3.Xiaoyao Decoction group can significantly reduce the visceral hypersensitivity of rats with liver depression and spleen deficiency IBS-D model,and is better than4.Pivectium bromide group in improving the symptoms and biological indicators of rats.5.Part of the mechanism of action of Xiaoyao decoction in the treatment of liver stagnation and spleen deficiency IBS-D may be related to VIP,CGRP,ZO-1,Claudin-4and Occludin indexes in colon tissue.6.Combined with experimental and clinical studies,it is confirmed that Xiaoyao Decoction is safe and effective in treating liver depression and spleen deficiency IBS-D,which has important clinical and theoretical value. |
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