| Functional Defecation Disorder(FDD)is a series of gastrointestinal diseases including the primary symptoms of continuing difficult defecation,reducing stool frequency or endless defecating sensation,accompanied by malfunction of bowel movements or abnormal pelvic floor muscles,and anxious or depressive emotions in most patients,which cause great stress on their mental state and lead to a vicious cycle of interaction between the body and mind.In recent years,with the development of psychosomatic medicine and the increasing stressful life,FDD has attracted more and more attention in clinical researches.However,there have been few studies on FDD pathogenesis,consequently,it is difficult to carry on the other related researches.Biofeedback therapy(BFT),the standard treatment of FDD,has achieved a good clinical efficacy.While,in the clinical practice,we found that some patients with biofeedback therapy only could not get a satisfactory effect,and some of them even showed tolerance to treatment,recurrence and relapse.Acupoint-embedding method in Traditional Chinese Medicine(TCM),based on acupuncture theories,is a characteristic TCM therapy,belonging to the external therapy of TCM.It is effective for many functional gastrointestinal diseases in written records and clinical practice.But we need modern research to prove its efficacy and probe its mechanism in FDD treatment.This study aimed to include the following aspects:1.to observe the clinical efficacy of the acupoint-embedding therapy by clinical research;2.to establish FDD animal model meeting Rome Ⅳ diagnostic standard,and explore its pathogenesis from the aspects of inflammation,brain-gut peptites,gastrointestinal motility,and ethology by animal experiments;3.to research the treatment of acupoint-embedding therapy for FDD and its mechanism from the aspects of inflammation,brain-gut peptides,gastrointestinal motility,and ethology.Clinical Research section:Clinical observation on acupoints embedding combined with biofeedback for functional defecation disorder.Objective:To observe the clinical effect of AET combined with BFT on patients with FDD and the effect on psychological status.Methods:We enrolled patients diagnosed with functional defecation disorder(FDD)in the Anorectal Clinic,Oriental Hospital of Beijing University of Chinese Medicine,from March 2018 to May 2019.Sixty FDD patients were randomly divided into a treatment group(30 cases)and a control group(n=30)according to the random number table method.Both groups received biofeedback treatment.The treatment group was treated with acupoints embedding based on biofeedback therapy.The treatment course was 8 weeks.The clinical effectiveness,constipation score before and after treatment,constipation quality of life questionnaire(PAC-QOL),resting anal canal pressure(ARP),anal canal systolic pressure(AMCP),and anal defecatory pressure(ADP)and duration of contraction of external anal sphincter(ALCT)were observed and recorded.Self-rating anxiety scale(SAS)and self-rating depression scale(SDS)were used in analysis.Results:The total clinical efficiency of the treatment group(92.86%)was significantly higher than that of the control group(69.23%)(P<0.05).After treatment,the constipation symptom score and ADP were significantly lower than before,and ALCT was significantly increased(P<0.05).After treatment,the feeling of defecation,time of defecation,falling,insufficiency,bloating and abdominal distension,scores of PAC-QOL,SAS,SDS and SAS of the control group were significantly lower than those before the treatment(P<0.05).After treatment,the PAC-QOL score,SDS score,AMCP,and ADP of the treatment group were significantly lower than those of the control group,while the ALCT score was significantly increased(P<0.05).Conclusion:Acupoints embedding,combined with biofeedback therapy,can more effectively improve the clinical efficacy of clinical symptoms,anorectal pressure,and psychological state in FDD patients.This combination is worthy of clinical promotion,since the efficacy is not satisfied with biofeedback therapy only in some cases.Animal Experiment section:Part 1:A rat model of functional defecation disorder simulated by low-fiber diet combined with perianal injection of methylene blue—a model evaluation.Objective:To establish a reasonable FDD modeling method by combination of a low-fiber diet and different doses of methylene blue perianal injection,and explore the pathogenesisMethods:36 male SD rats were randomly divided into blank group(BG),low-fiber diet group(LFDG),lidocaine group(LG),low dose of methylene blue group(MBG-L),medium dose of methylene blue group(MBG-M),and high dose of methylene blue group(MBG-H).The blank group was fed up by ordinary breed feed,and the rest of other groups were by low-fiber diet feeding.In 9th week,lidocaine and methylene blue(low,medium,high)groups got an perianal injection with 2%lidocaine,0.05%methylene blue,0.1%methylene blue,and 0.2%methylene blue,about 1-2ml,respectively.(Methylene blue injection was a mixture,which was made up of 1%methylene blue injection and 2%lidocaine injection by concentration.)In 14 days,we observed weight changes,feces,behavioral changes,and tissue changes of the colon,rectum and brain;detected 5-HT,IL-6,TNF alpha,substance P and concentration of Ach in the serum,and ACHE and CGRP levels in the tissues;tested location and expression quantity of CGRP protein by using Western Blot(WB),and immunohistochemical method(IHC)in the rectum tissue;and measured CGRP mRNA expression by using real-time fluorescent quantitative PCR method in the tissues of rats.Results:During the modeling period,the fecal weight of rats in MBG-(L,M,H)was significantly lower than that of BP,the difference was statistically significant(P<0.05),and the fecal quality of MBG-M was the lowest among those groups.In the large intestine dynamics,the extension time of MBG-(L,M,H)in analogue balloon expulsion test in 9th and 10th weeks was obviously higher than that of BG,LFDG and LG,and the difference was statistically significant(P<0.05).There was no statistically significant difference among MBG-L to MBG-H(P>0.05).The declining anorectal resting pressure in 9th and 10th weeks of MBG-(L,M,H)was significantly lower than that of BG,LFDG and LG,with statistically significant difference(P<0.05),in which MBG-M was the lowest group compared to MBG-L and MBG-H(P<0.05).There was no significant difference among the six groups in intestinal movement test by using charcoal powder(P>0.05).In terms of behavioral test,there was a slight decrease in sugar-water preference test in the modeling group,with no significant difference(P>0.05).In 9th and 10th weeks after the simulation,MBG-(L,M,H)showed a significantly lower movement in open field experiment of horizontal movement,vertical movement,and the central cell residence time,as well as a decrease of time percentage and frequency percentage of opening arm entrance in overhead cross maze experiment,which are compared to those of BG and LFDG,with statistically significant difference(P<0.05).There was no significant difference among MBG-L、MBG-M and MBG-H(P>0.05).In serological test,the amount of 5-HT,substance P,and Ach in the serum of MBG-L、MBG-M and MBG-H was significantly lower than that of BG,LFDG and LG，and the difference was statistically significant(P<0.05).Among them,MBG-M was the lowest group compared with MBG-L and MBG-H,and the difference was statistically significant(P<0.05).Meanwhile,the serum content of IL-6 and TNF alpha in MBG-L、MBG-M and MBG-H was significantly higher than that in BG,LFDG and LG,and the difference was statistically significant(P<0.05).Among them the MBG-M got the highest content compared to MBG-L and MBG-H(P<0.05).In brain,colon and rectum tissue,the content of CGRP and ACHE in MBG-L、MBG-M and MBG-H was higher than in the BG and the LFDG,which got a statistically significant difference(P<0.05).The content of CGRP in brain,colon tissues of MBG-M was lower than in MBG-L and MBG-H(P<0.05).The CGRP protein in the rectum of the MBG-L、MBG-M and MBG-H were decreased by Western Blot detection,and a significant decrease was found in the MBG-M and MBG-H,with statistically significant difference from the BG(P<0.05).The integrated optical density(IOD)of CGRP protein in colon and rectal tissues of the MBG-L、MBG-M and MBG-H was decreased by IHC method,among which MBG-M and MBG-H were significantly higher than the BG,the LFDG and LG,with statistically significant difference(P<0.05).The CGRP mRNA expression in colon and rectum of the MBG-L、MBG-M and MBG-H was significantly higher than that of the BG,the LFDG by Real time PCR(P<0.05),and the difference was statistically significant(P<0.05).The CGRP mRNA expression in the brain tissues of the MBG-L、MBG-M and MBG-H was slightly higher than that of the BG,the LFDG,and the difference between the groups was not statistically significant(P>0.05).Conclusions:1.The low-fiber diet combined with methylene blue perianal injection is a good way to simulate FDD,such as fecal character,anal dynamics and ethology,etc.This model is worth extending in research.2.The pathogenesis of FDD may be related to inflammation and disorder of intestinal nerve regulation.However,it is still necessary to further study whether the two reasons causing diseases singly or in combination.3.Based on the comprehensive evaluation of fecal characteristics,anal dynamics,ethology,histopathology and serology,and considering safety,economic and environmental protection factors,the low-fiber diet combined with methylene blue medium dose perianal injection is recommended for establishing research model.Part 2:A experimental Study on Mechanism of Acupoints Embedding in Intervening Functional Defecation Disorder Model in RatsObjective:To observe the intervention of acupoints-embedding therapy for FDD model rats and analyze its mechanism by experimental studyMethods:Twenty-four male SD rats were randomly divided into a blank group,a model group,an acupoint-embedding group,and a sham intervention group.Rats in the blank group were fed with a regular diet.The remaining three groups were modeled according to the "low fiber diet combined with 0.1%methylene blue perianal injection(methylene blue medium dose)method" mentioned in experiment 1.At the 9th week(the day after the perianal injection of methylene blue)after modeling:1.The acupoint-embedding group was treated with acupoint-embedding.2.The sham treatment group was treated with acupoint acupuncture.During the treatment,we observed the changes in body weight,feces,defecation patterns,and behavior of rats.After the treatment,we observed the general tissue changes in the colon,rectum,and brain tissue of each group of rats.At the same time,the rats of each group were tested:1.The concentrations of 5-HT,IL-6,TNF-α,substance P,Ach in serum;2.Detection of CGRP protein localization and expression in rat rectal tissues by Western Blot(WB)and immunohistochemistry(IHC);3.The expression of CGRP mRNA in rat tissues was measured by real-time fluorescent quantitative PCR.Result:Before the intervention,the weight of rats at the model group,acupoint-embedding group,and sham treatment group was significantly lower than that of the blank group,and the difference was statistically significant(P<0.05).Among them,the weight of rats at the acupoint-embedding group was significantly lower than that of the model group and the sham treatment group,and the difference was statistically significant(P<0.05).In the first and second week after the intervention,the weight of rats at the acupoint-embedding group,and sham treatment group was significantly lower than that of the blank group,and the difference was statistically significant(P<0.05).Among them,the weight of rats at the acupoint-embedding group was significantly lower than that of the model group and the sham treatment group,but the difference was not statistically significant(P>0.05).Before and in the first week and the second week after the intervention,the stool quality of the rats in the model group,the acupoint-embedding group and the sham treatment group was lower than that of the rats in the blank group,and the difference was statistically significant(P<0.05).In the first week and the second week after the intervention,the stool quality in the acupoint-embedding group and the sham treatment group was significantly higher than that in the model group,and the difference was statistically significant(P<0.05).Colorectal dynamics:There was no significant difference in the simulated balloon discharge time among the groups before the intervention(P>0.05);after the first and second week of intervention,the balloon discharge time of the acupoint-embedding group was significantly higher than that of the blank group but significantly lower than that of the model group and the sham treatment group,and the difference was statistically significant(P<0.05).After 1 and 2 weeks of intervention,the anorectal resting pressure of the rats in the acupoint-embedding group was significantly higher than that of the model group and the sham-treated group,and significantly lower than that of the blank group,and the difference was statistically significant(P<0.05).There was no significant difference among all the groups in the charcoal propelling experiment(P>0.05).In terms of animal behavior,after two weeks’ intervention,the syrup preference in the acupoint-embedding group was significantly higher than that in the model group,and the difference was statistically significant(P<0.05).In the open field experiment,the rats in the acupoint-embedding group had higher horizontal movement,vertical movement,and longer central grid staying time than the rats in the model group and the sham treatment group,but lower than those in the blank group.The horizontal movement of rats in the acupoint-embedding group was significantly higher than that in the other groups after one week of intervention,and the difference was statistically significant(P<0.05).One week after the intervention in the acupoint-embedding group,the percentage of time to enter the open arm was significantly higher than that in the model group and the sham treatment group,and the difference was statistically significant(P<0.05).Rats in the acupoint-embedding group had significantly higher arm opening time and frequency than those in the model group and the sham treatment group in two weeks after intervention,andthe difference was statistically significant(P<0.05).In terms of serological evaluation,5-HT,substance P,and Ach in rats in the acupoint-embedding group were significantly higher than those in the model group and the sham treatment group,but at the same time significantly lower than those in the blank group,and the differences were statistically significant(P<0.05).IL-6 and TNF-α in the acupoint-embedding group were significantly lower than those in the model group and sham treatment group,but at the same time significantly higher than those in the blank group,and the differences were statistically significant(P<0.05).By using the WB method,the content of CGRP in the rectum and colon of rats in the acupoint-embedding group was lower than that in the model group and sham treatment group,but at the same time higher than that in the blank group,and there was no significant difference among the groups(P>0.05).With the IHC method,the value of the integrated optical density of CGRP protein in the colon and rectum of the acupoint-embedding group rats was significantly lower than that of the model group rats,but at the same time significantly lower than that of the blank group rats,and the differences were statistically significant(P<0.05).The expression of CGRP mRNA in the rectum and colon of rats in the acupoint-embedded group detected by RT-PCR was significantly lower than that in the model group and the sham-treated group but at the same time significantly higher than that in the blank group.The difference is statistically significant(P<0.05).Conclusions:Acupoint-embedding therapy can effectively improve the symptoms of constipation,anorectal dynamics and behavioral performance in FDD rats.The mechanism may be related to gastrointestinal motility,improved inflammatory status,and CGRP protein changes.Nevertheless,its correlation with the brain-gut pathway needs further research. |
PDF Full Text Request