| Objective: Under the guidance of "large intestine" theory,the animal model of slow transmission constipation was replicated,the changes of function and expression of AQP3 and AQP9,the activity level of the regulating factors of intestinal neurotransmitters and inflammatory factors were observed,and the dry preaction of the "Zeng liquid boat" method was given by the Decoction of Zeng liquid,through its "dose effect",The "time effect" relationship and "square test" confirm that AQP3 and AQP9 mediated disturbance of the colon water metabolism and control failure are one of the important links in the pathogenesis of STC,in order to find a new theory of STC,to treat new targets,and to provide reference for the research and development of cathartic drugs,and to provide a basis for the study of the mechanism of the catharsis of traditional Chinese medicine.Materials and methods:Part one: clinical studyClinical study on the treatment of slow transit constipation patients with Zeng liquid DecoctionFrom March 2016 to March 2017 were consistent with the diagnosis of slow transit constipation in 80 patients with anorectal clinic patients combined with traditional Chinese medicine and Western medicine in Affiliated Hospital of Chuanbei Medical College,were randomly divided into two groups,control group and treatment group respectively,40 cases in each group,the control group was given oral Marenrunchang pills treatment,the treatment group was given.Liquid decoction,treatment for two weeks,the two groups of patients were followed up,follow-up include: defecation of patients before and after treatment,such as defecation time,frequency and stool score,symptom assessment,and the presence of abdominal pain and distension and other gastrointestinal symptoms,and make detailed records,according to the 2012 constipation disease(constipation colonic)of TCM diagnosis and treatment plan,the statistics of the treatment group and control group before and after treatment,the main symptom scores,and calculate the total score.The treatment effect of two groups of patients before and after medication and the difference of the curative effect of two kinds of drugs were analyzed.The second part: basic researchThe first chapter: the establishment of the rat model of slow transit constipationThe application of Compound Diphenoxylate Tablets copy "liquid deficiency" model of slow transit constipation: Compound Diphenoxylate Tablets according to human and animal drug is converted into the corresponding dose of 10mg/kg plus saline 4ml at a concentration of suspension of 50%,at the same time except blank control group rats were given limited water treatment,namely normal drinking water.1/3,the integer 10 ml,given 4 times by gavage,in addition to the first Compound Diphenoxylate Tablets 3ml suspension,the rest are 2ml,3ml,2ml,ultra pure water,irrigation time respectively is: 8:00,12:00,16:00,20:00,by gavage for 7 days.The model was successfully replicated by comparing the general condition of the rat and the experimental model of the propelling experiment.After the success of the model,the blank group and the model group were killed.The second chapter: the effect of Zeng Ye Decoction on the distribution and expression of AQP3 and APQ9 in the colon of slow transit constipation rats120 SD rats were randomly divided into 12 groups: control group,model group,itopride group,Zengye Decoction low dose group,dose group by liquid soup,Zengye Decoction high dose group,Zengye Decoction group for 24 hours,48 hours of Zengyetang group,Zengye Decoction of 72 hours of Zengyetang group,96 hours group,Zengyetang group for 120 hours,144 hours of Zengyetang group,10 rats in each group,half male and half female.Except the normal group,the other groups were given by gavage diphenoxylate induced "liquid deficiency" slow transit constipation rat model,after the success of modeling,the rats were given corresponding drugs by gavage treatment,observation of Zengyetang on slow transit constipation rats laxative effect,detected by immunohistochemistry group AQP3,AQP9 and expression in colon and the distribution of software to automatically select all areas with special color image analysis,determination of the average optical density in the region(MOD)value,and ultimately mean on behalf of AQP3 and AQP9 positive expression of AQP3 and AQP9 parameters;detected by Q-PCR in colonic tissue level,protein immunoblotting(Western blot)to detect AQP3 and AQP9 protein expression.The third chapter: the effect of Zeng Ye Decoction on the activity of VIP,SP,5-HT,NO,Ach,PGE2 and TNF-alpha in the serum of slow transit constipation ratsThe group was divided into second chapters.The rats in the experimental group were sacrificed after the corresponding intervention.The blood sampling method wastaken before the death of the abdominal aorta.The rats were killed by the dislocated of the cervical vertebra after the blood was taken.The blood samples were stored by EP tube,30 min was stored in the tube,and the plasma was centrifuged at 4 degrees C.The supernatant was separated into the EP tube after centrifugation,and the refrigerator was preserved at-20 C.The detection of cytokine VIP,SP,5-HT,NO,Ach,PGE2 and TNF-alpha in serum was performed strictly according to the procedures of ELISA kit.Result:Clinical study of Zeng Ye Tang in treating slow transit constipation due to "liquid deficiency"1,The treatment group and the control group were no significant difference in general,such as gender,age,course of disease was comparable,no statistical significance(P>0.05),treatment group and control group before and after treatment the total score had significant difference(P<0.01)between the two groups of patients after treatment,the total score have significant difference(P<0.01).The effective rate of the treatment group was 78.1%,and the effective rate of the control group was 71%,indicating that the clinical efficacy of the two drugs after treatment was statistically significant(P<0.05),and the curative effect of the treatment group was better than that of the control group.2,the establishment of a rat model of slow transit constipationCompared with the blank group,the faeces of the rats in the modeling group changed from dry wet yellow strip to dry and hard brown granules,and their defecation volume was reduced,mental activity and activity were significantly worse than before,and their hair glossiness and weight gain increased slowly.After modeling the rats,the excretion time of the first feces and black stools slowly slowed down,and the fecal water content of the stool was 24 hours and 24 hours.The difference was statistically significant(P <0.01).It is suggested that the model is successful.With the blank group(carbon ink propulsion rate was 64.65%)compared to the model group(ink advance rate of 56.28%)carbon ink rat advance rate decreased significantly,revealed a statistically significant difference(P=0.05),it is indicated that the application of compound diphenoxylate can make rat colonic peristalsis slows down,fecal transit time prolonged,indicating slow transit constipation model of success.3,The effect of Zeng Ye Decoction on the distribution and expression of AQP3 and APQ9 in the colon of slow transit constipation ratsCompared with the blank group,the content of AQP3 in the proximal colonic tissue of the model group increased significantly(P < 0.05),and the content of AQP9 in the distal colon tissue decreased significantly(P < 0.05).After the treatment of the decoction and the treatment of the model group,the proximal colon AQP3 of the high dose group,the IDP,and the 72 hour group of the decoction of Zeng liquid was compared with the model group.The content of the distal colon AQP9 increased significantly(P < 0.05),and the content of AQP9 in the distal colon increased significantly(P < 0.05),and the difference was statistically significant.The content of the proximal colon in the low dose group and the 48 hour group of Zeng liquid Decoction decreased,and the content of AQP9 in the distal colon group increased,but the difference was not significant(P>0.05),and the decoction of Zeng liquid decoction was 24 hours.In the proximal colonic tissue,there was no significant decrease in AQP3(P>0.05),and there was no significant increase in the content of AQP9 in the distal colon(P>0.05).The content of AQP3 in the proximal colon of the96,120,and 144 hour group decreased significantly(P < 0.01),and the content of AQP9 in the distal colon was significantly increased(P < 0.01).The difference was statistically significant.The difference was significant,and there was no difference between the three groups.4,the effect of Zeng Ye Decoction on the activity of VIP,SP,5-HT,NO,Ach,PGE2 and TNF-alpha in the serum of slow transit constipation ratsThe serum concentration of VIP and PGE2 in the serum of ELISA test group was significantly higher than that in the blank group.The difference was statistically significant(P < 0.05).The concentration of VIP in the group of Yi Tong Li,the decoction of Zeng liquid,the high dose group,the 96,120 and 144 hour group of the decoction of Zeng liquid was lower than that of the model group,and the difference was statistically significant.The low dose group and the decoction of Zeng liquid Decoction 24,48 and 72 were significant.The serum VIP concentration in hourly group was not significantly lower than that in model group(P>0.05).The concentration of SP in the serum of the model group was significantly lower than that in the blank group.The difference was statistically significant(P < 0.05).The concentration of SP in the group,the high dose group,the 72,96,120 and 144 hour group of the treatment group was higher than that of the model group,and thedifference was statistically significant.The serum concentration of the decoction,the low dose group,the 24 and 48 hour group of the decoction of Zeng liquid decoction was SP.Compared with the model group,the concentration increased,but the difference was not statistically significant(P>0.05).The concentration of 5-HT in the serum of the model group was significantly higher than that in the blank group,and the difference was statistically significant(P <0.05).The concentration of 5-HT in the group of Yi Tong Li,the decoction of Zeng liquid,the high dose group,the 72,96,120 and 144 hour group decreased,and the difference was statistically significant(P < 0.05).The low dose group of Zeng decoction,the soup 24 and 48 were small.There was no significant difference in serum 5-HT concentration between the two groups(P>0.05).The concentration of NOS in the serum of the model group was significantly higher than that in the blank group.The difference was statistically significant(P <0.05).The concentration of NOS in the group,the high dose group,the 96,120 and144 hour group of the treatment group decreased,and the difference was statistically significant(P < 0.05).The concentration of the decoction,the low dose group,the decoction of Zeng liquid,24,48,72 were small.There was no significant difference in serum NOS concentration between the two groups(P>0.05).The concentration of Ach in the serum of the model group was significantly lower than that in the blank group.The difference was statistically significant(P <0.05).The concentration of Ach in the group,the high dose group,the 120 and 144 hour group increased in the model group,and the difference was statistically significant(P < 0.05).The low,middle dose group and the decoction of Zeng liquid Decoction were 24,48,72 and 96.The concentration of Ach in the serum of the time group was higher than that in the model group,but the difference was not statistically significant(P>0.05).The concentration of TNF-alpha in the serum of the model group was higher than that in the blank group,and there was no statistical significance(P>0.05).There was no significant change in the other experimental groups compared with the blank group,and there was no statistical significance(P>0.05).Conclusion:1,a common clinical "liquid deficiency" in patients with slow transit constipation,increase curative effect of compound Chinese medicine decoction clinically significant,compared with the better effect of Maren Runchang pills.2,the application of compound diphenoxylate copy "liquid deficiency" slow transit constipation rat model with disease formation process,can be copied successfully.3,the proximal AQP9 increased the content of AQP3 in colon tissue and distal colon tissues decreased may be one of the pathogenesis of STC;Zengyetang through down-regulation of AQP3 expression and upregulate the expression of AQP9,may play the "liquid Zhou" effect mechanism and treatment of STC.4,The change of serum VIP,SP,5-HT,NO,Ach,PGE2,TNF-alpha activity level may be one of the pathogenesis of Tianjin loss of slow transit constipation,Zengyetang by regulating various intestinal neurotransmitters,inflammatory factor expression to treat "liquid deficiency" slow transit constipation,may is the mechanism to play a "fluid increasing effect of Zhou". |
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