Study On The Mechanism Of Simo Decoction In The Treatment Of Functional Constipation Based On The Theory Of Brain-Gut Axis

Objective1 Observe the therapeutic effect of Simo Decoction(SMD)on clinical symptoms and quality of life in functional constipation(FC)patients with Intestinal Qi Stagnation;evaluate the improving effect of SMD on physical health and mental health;explore the correlation between the SMD clinical effect and the degree of improvement of life quality.2 Establish gastrointestinal hypomotility animal models with intestinal qi stagnation being used of psychological stress,gastrointestinal stimulation factors.Observe the changes eneral status,gastrointestinal motility,psychological and behavioral changes of animal models before and after the treatment of SMD;detect the expression of brain-gut peptide in central nervous system(CNS),enteric nervous system(ENS);observe the similarities and differences of expression in myenteric plexus before and after treatment of SMD.Method1 Forty-five FC patients with Intestinal Qi Stagnation were selected at the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine During October 2012 to April 2013.Randomly 30 patients were given SMD treatment and 15 patients were given mosapride and deanxit for 2 weeks.Before and after treatment,Chinese medicine questionnaire were answered by patients to evaluate the general condition,the main and associated symptoms,behavior of life,;use short Form 36 questionnaire(SF-36)to evaluated the quality of life(physical health and mental health)of patients.2 108 SPF Sprague Dawley(SD)rats were selected,female,weight about 160-180g,adaptive feeding for 1 weeks.Then all rats were randomly divided into 9 groups:normal group,psychological stress model group(control,low dose,high dose and mosapride group)and gastrointestinal stimulation model group(control,low dose,high dose and mosapride group),12 rats in each group.Using tail clamping and immobilization to establish psychological stress model;using Rhubarb and ice water administration to establish gastrointestinal stimulation model,the modeling time is 35 days.Normal group were fed with normal diet,water supply;model group were given 2ml saline intragastric orally;1.8g/kg,3.6g/kg SMD were administered to SMD low dose group(LDG)and SMD high dose group(HDG);1.5mg/kg mosapride were administered to mosapride group for seven days.Observe the general status of the rats before modeling,after modeling,and after SMD treatment;evaluated the score of test of Open-Field and Sucrose-Consumption before modeling and after SMD treatment;detecte the rates of gastric emptying and intestinal propulsion;detecte the expression of brain-gut peptide Substance P(SP),Somatostatin(SS)in hypothalamus and intestinal myenteric plexus;and detect the expression of Protein Gene Product 9.5(PGP9.5)in intestinal myenteric plexus.Results1 clinical research1.1 General InformationIn SMD group,there were 12 males,18 females,aged from 25 to 77 years(51.67±15.10 years),clinical course from 8 months to 13 years(6.59±4.04 years).In western medicine group,there were 5 males,10 females,aged from 37 to 73 years(52.40 ± 12.29 years),clinical course from 2 to 10 years(5.20±2.65 years).There were no statistical significance between SMD group and western medicine group in gender,age,and course of disease(P>0.05),two groups have comparability.1.2 Symptom score1.2.1 After SMD treatment,main symptoms in FC patients with Intestinal Qi Stagnation were significantly lower in the scores of defecation frequents,defecation time,defecation difficulty,un-comfort/falling/swelling and distention symptom compared with those before treatment(P<0.05);there is no statistical significance in the scores of stool scale(P>0.05).1.2.2 After SMD treatment,associated symptoms in FC patients with Intestinal Qi Stagnation were significantly lower in the scores of abdominal pain,early satiety,belching appetite,irritability,depressed and insomnia symptoms compared with those before treatment(P<0.05);there is no statistical significance in the scores of bowel and fart(P>0.05).1.2.3 After SMD treatment,the average scores of main symptoms,associated symptoms and total symptoms were significantly lower than those before treatment(P<0.05).1.3 Clinical curative effectThere are no statistically difference in main symptoms,accompany symptoms and total symptoms between SMD group and western medicine group(P>0.05,P>0.05,P>0.05).1.4 Assessment of life qualityAfter SMD treatment,life quality in FC patients with Intestinal Qi Stagnation were significantly higher in the scores of Physiological Function(PF),Role Physical(RP),Bodily Pain(BP),General Health(GH),Vitality(VT),Social Function(SF),Role Emotional(RE),Mental Health(MH)than those before treatment(P<0.05).Life quality in FC patients with Intestinal Qi Stagnation were significantly higher in the scores of Physical Component Summary(PCS)and Mental Component Summary(MCS)than before.1.5 Correlation between improvement of life quality and clinical curative effect.There were significantly positive correlation between main symptoms curative effect and PF(P<0.05),GH(P<0.05),VT(P<0.01)and MH(P<0.01);there were significantly positive correlation between accompany symptoms curative effect and PF(P<0.05),RP(P<0.05),RE(P<0.05),GH(P<0.01),VT(P<0.01),MH(P<0.01);there were significantly positive correlation between total symptoms curative effect and PF(P<0.05),RP(P<0.05),SF(P<0.05),RE(P<0.05),GH(P<0.01),VT(P<0.01),MH(P<0.01).There were significantly positive correlation between main,accompany and total symptoms curative effect and PCS(P<0.05),MCS(P<0.01),and MCS has more relevance than PCS.2 Experimental studies2.1 General stateThe rats in normal group,fur shiny,spiritual,lively and playful,eating normally.Psychological stress group rats presentation of restlessness,irritability,constantly screaming at begging,and activities reduced,disturbed easily,perform horripilation at later.Gastrointestinal stimulation group rats apathetic,sleepy and somnolence,fur lackluster,reaction slowed down,food intake and water consumption reduced.After SMD treatment,rats activities increase,appetite increase,sensitive reaction,fur shiny.2.2 Weight changeBefore modeling(Day 0):there is no significant difference between each group(P>0.05).After modeling(D35):compared with normal group,the body weight in each group of psychological stress and gastrointestinal stimulation decreased significantly(P<0.05);compared with model group,there are no significant difference in the body weight in all treatment groups,each group has comparability.After SMD treatment(D42):compared with normal group,the body weight in psychological stress model group and gastrointestinal stimulation model group decreased significantly(P<0.05),and the data in gastrointestinal stimulation model group decreased more significantly.Compared with gastrointestinal stimulation model group,the body weight of LDG,HDG,and mosapride group increased significantly(P<0.05);compared with psychological stress model group,the body weight of LDG,HDG,and mosapride group increased significantly(P>0.05).2.3 Gastrointestinal motility2.3.1 Gastric emptying rateCompared with normal group,the rate of gastric emptying in psychological stress model group and gastrointestinal stimulation model group decreased significantly(P<0.05).Compared with gastrointestinal stimulation model group,the rate of gastric emptying in LDG,HDG,and mosapride group increased(P>0.05,P<0.05,P>0.05);compared with psychological stress model group,the rate of gastric emptying in LDG,HDG,and mosapride group increased(P>0.05,P<0.05,P<0.05).2.3.2 Bowel propulsion rateCompared with normal group,the rate of bowel propulsion in psychological stress model group and gastrointestinal stimulation model group decreased significantly(P<0.05).Compared with gastrointestinal stimulation model group,the rate of bowel propulsion in LDG,HDG,and mosapride group increased(P>0.05,P<0.05,P>0.05);compared with psychological stress model group,the rate of bowel propulsion in LDG,HDG,and mosapride group increased(P>0.05,P<0.05,P<0.05).2.4 Psychological behavior2.4.1 Open-Field test2.4.1.1 Horizontal Movement:Compared with normal group,the score of horizontal movement in psychological stress model group and gastrointestinal stimulation model group decreased significantly(P<0.05),and the data in gastrointestinal stimulation model group decreased more significantly.Compared with gastrointestinal stimulation model group,the score of horizontal movement in LDG,HDG,and mosapride group increased(P>0.05,P>0.05,P<0.05);compared with psychological stress model group,the score of horizontal movement in LDG,HDG,and mosapride group increased(P>0.05,P<0.05,P<0.05).2.4.1.2 Vertical Movement:Compared with normal group,the score of vertical movement in psychological stress model group and gastrointestinal stimulation model group decreased significantly(P<0.05),and the data in psychological stress model group decreased more significantly.Compared with gastrointestinal stimulation model group,the score of vertical movement in LDG,HDG,and mosapride group increased(P>0.05,P>0.05,P<0.05);compared with psychological stress model group,the score of vertical movement in LDG,HDG,and mosapride group increased(P>0.05,P>0.05,P<0.05).2.4.2 Sugar-Consumption testCompared with normal group,the consumption of sucrose and consumption rate of sucrose in psychological stress model group and gastrointestinal stimulation model group decreased obviously(P<0.05),and the data in gastrointestinal stimulation model group decreased more significantly.Compared with gastrointestinal stimulation model group,the consumption of sucrose and consumption rate of sucrose in LDG,HDG,and mosapride group increased(P>0.05,P<0.05,P>0.05);compared with psychological stress model group,the consumption of sucrose and consumption rate of sucrose in LDG,HDG,and mosapride group increased(P>0.05,P<0.05,P<0.05).2.5 Expression of brain-gut peptide in intestinal myenteric plexus2.5.1 Expression of Substance P in intestinal myenteric plexusUnder microscope,expression of Substance P in intestinal myenteric plexus were brown color expression,mainly exist in cytoplasm,and nucleus are blue color.2.5.1.1 Spectroscopic analysis on Substance P in intestinal myenteric plexusGastrointestinal stimulation model group:there is statistical significance between control group and normal group(P<0.05);there are statistical significance between LDG,HDG,mosapride group and control group(P<0.05).Psychological stress model group:there is no statistical significance between control group and normal group(P>0.05);there are no statistical significance between LDG,HDG,mosapride group and control group(P>0.05).2.5.1.2 Mean Density on Substance P in intestinal myenteric plexusCompared with normal group,mean density on Substance P in intestinal myenteric plexus in gastrointestinal stimulation model group and psychological stress model group decreased significantly(P<0.05,P<0.05)).Compared with gastrointestinal stimulation model group,mean density on Substance P in intestinal myenteric plexus in LDG,HDG,and mosapride group increased(P<0.05,P<0.05,P<0.05);compared with psychological stress model group,mean density on Substance P in intestinal myenteric plexus in LDG,HDG,and mosapride group increased(P>0.05,P>0.05,P<0.05).2.5.2 Expression of Somatostat in intestinal myenteric plexusUnder microscope,expression of Somatostat in intestinal myenteric plexus were brown color expression,mainly exist in cytoplasm,and nucleus are blue color.2.5.2.1 Spectroscopic analysis on Somatostat in intestinal myenteric plexusGastrointestinal stimulation model group:there is statistical significance between control group and normal group(P<0.05);there are statistical significance between LDG,HDG,mosapride group and control group(P<0.05).Psychological stress model group:there is no statistical significance between control group and normal group(P>0.05);there are no statistical significance between LDG,HDG,mosapride group and control group(P>0.05).2.5.2.2 Mean Density on Somatostat in intestinal myenteric plexusCompared with normal group,mean density on Substance P in intestinal myenteric plexus in gastrointestinal stimulation model group decreased significantly(P<0.05),and the data in psychological stress model group increased(P<0.05).Compared with gastrointestinal stimulation model group,mean density on Somatostat in intestinal myenteric plexus in LDG,HDG,and mosapride group increased(P>0.05,P>0.05,P>0.05);compared with psychological stress model group,mean density on Substance P in intestinal myenteric plexus in LDG,HDG,and mosapride group decreased(P>0.05,P<0.05,P<0.05).2.6 Expression of brain-gut peptide in hypothalamusUnder microscope,expression of Substance P and Somatostat in hypothalamus were brown color expression,mainly exist in cytoplasm and membrane,and nucleus are blue color.2.6.1 Mean Density on Substance P in hypothalamusCompared with normal group,mean density on Substance P in hypothalamus in gastrointestinal stimulation model group increased(P<0.05),and the data in psychological stress model group decreased(P<0.05).Compared with gastrointestinal stimulation model group,mean density on Somatostat in hypothalamus in LDG,HDG,and mosapride group decreased(P<0.05,P<0.05,P<0.05);compared with psychological stress model group,mean density on Substance P in hypothalamus in LDG,HDG,and mosapride group increased(P<0.05,P<0.05,P<0.05).2.6.2 Mean Density on Somatostat in hypothalamusCompared with normal group,mean density on Somatostat in hypothalamus in gastrointestinal stimulation model group decreased(P<0.05),and the data in psychological stress model group increased(P<0.05).Compared with gastrointestinal stimulation model group,mean density on Somatostat in hypothalamus in LDG,HDG,and mosapride group increased(P>0.05,P<0.05,P<0.05);compared with psychological stress model group,mean density on Somatostat in hypothalamus in LDG,HDG and mosapride decreased(P>0.05,P<0.05,P<0.05).2.7 Expression of Protein Gene Product 9.5 in intestinal myenteric plexus2.7.1 Pathological observationUnder microscope,expression of Protein Gene Product 9.5 in intestinal myenteric plexus were brown color expression,mainly exist in cytoplasm.In gastrointestinal stimulation model group,muscle fiber structure disorder or even disconnected and nerve cells decreased.In gastrointestinal stimulation model group are affected to a lesser degree.After SMD treatment,muscle fiber structure and the number of nerve cells recovery more or less.2.7.2 Spectroscopic analysis on Protein Gene Product 9.5 in intestinal myenteric plexusGastrointestinal stimulation model group:there is statistical significance between control group and normal group(P<0.05);there are statistical significance between HDG and mosapride group and control group(P<0.05),there is no statistical significance between LDG and control group(P>0.05).Psychological stress model group:there is no statistical significance between control group and normal group(P>0.05);there are no statistical significance between LDG,HDG,mosapride group and control group(P>0.05).2.7.3 Mean Density on Protein Gene Product 9.5 in intestinal myenteric plexusCompared with normal group,mean density on Protein Gene Product 9.5 in intestinal mventeric plexus in gastrointestinal stimulation model group and psychological stress model group decreased(P<0.05,P<0.05).Compared with gastrointestinal stimulation model group,mean density on Protein Gene Product 9.5 in intestinal myenteric plexus in LDG,HDG,and mosapride group increased(P<0.05,P<0.05,P<0.05);compared with psychological stress model group,mean density on Protein Gene Product 9.5 in intestinal myenteric plexus in LDG,HDG,and mosapride group increased(P>0.05,P>0.05,P>0.05).Conclusion1 Simo Decoction can alleviate main symptoms,associated symptoms,and improve quality of life in functional constipation(FC)patients with Intestinal Qi Stagnation2 Simo Decoction can promote bowel movement and improve psychological behavior in rats model with gastrointestinal stimulation and psychological stress.3 Simo Decoction can increase the expression of Substance P and Somatostat in intestinal myenteric plexus in gastrointestinal stimulation model group,it indicated that Simo Decoction can repair damaged intestinal mucosa and promote bowel movement;Simo Decoction can increase the expression of Substance P and reduce the expression of Somatostat in intestinal myenteric plexus in gastrointestinal stimulation model group,it indicated that Simo Decoction can promote bowel movement slowed down by psychological stress.Simo Decoction can reduce the expression of Substance P and increase the expression of Somatostat in hypothalamus in gastrointestinal stimulation model group,and reduce the expression of Substance P and Somatostat in hypothalamus in psychological stress model group,it indicated that Simo Decoction can adjust psychological behavior changes caused by gastrointestinal stimulation and psychological stress,and the function of brain-gut peptide are different in central nervous system(CNS)and enteric nervous system(ENS),ENS may regulate gastrointestinal physiology independently and CNE may respond to the change of ENS.4 Simo Decoction can increase the expression of Protein Gene Product 9.5 in intestinal myenteric plexus in gastrointestinal stimulation model group,it indicated that Simo Decoction can repair damaged intestinal mucosa caused by gastrointestinal stimulation.