Yang Rong The Intestines Shu Mixture Of Slow Transit Constipation A Therapeutic Effect And Mechanism Study

Constipation is a common clinical symptom.Changes in dietary structures and effects of psychosocial factors have been attributed as the cause of increased rates of constipation.It has been reported that the morbidity of chronic constipation is 6.07%in China and it has reaches 15-20%in aged population.Chronic constipation is a major predisposing factor for a variety diseases,such as colon cancer,hepatic encephalopathy,breast diseases and Alzheimers Disease.Furthermore,it can induce some life threatening diseases such as acute myocardial infarction and stroke.In order to develop effective approaches in constipation treatment,relief patients from the constipation and to reduce the incidence of severe diseases induced by constipation,it is important to make research into constipation an area of great importance.Slow transit constipation accounts for nearly half(approximately 45%) of cases in all kinds of constipation.Compared with western medicine,the advantages of traditional Chinese medicine in the prevention and treatment of slow transit constipation have been highlighted in recent years.More efficient effects and less adverse effects have been show in traditional Chinese medicine in treating slow transit constipation.YangRongRunChangShu(YRRCS) is a combined Chinese medicine which has been invented by Professor Tian Zhengguo.Based on large amount of clinical practice,and ancient and modern literatures,Prof.Tian has summarised YRRCS in treating slow transit constipation.For the constipation patients in Qi,Blood and Body Fluids Deficiency Type according to traditional Chinese differentiation,YRRCS can tonify qi and replenish blood and moisten the intestines.It can improve the function of colon by soothing the liver and regulating the spleen,tonifing the lung and reinforcing kidney,moistening the intestines and relax the bowels.The previous clinical study has shown the total effective rate of YRRCS is 92.5%.To provide the scientific evidence for the development of new Chinese medicine,the pharmacodynamics of YRRCS and its related mechanism have been investigated in this study.This study was divided into two parts.In the first part,effects of YRRCS on passing stools were examined in Experiment 1,2 and 3.In the second part,the mechanism of YRRCS on slow transit constipation treatment was investigated by using western blot method and immunohistochemistry method to examine the colon interstitial cells Cajal(ICC) and vasoactive intestinal peptide(VIP) and substance P(SP) in colon myenteric plexus of slow transit constipation mice model.Section one:The Study of Effects of YRRCSExperiment 1:Duplicate model of slow transit constipationObjective:Duplicate mouse model of slow transit constipationMethod:We chose 96 healthy mice and randomly divided them into six groups,i.e.normal group,model group,Mazirenwan group,slight-dose YRRCS group,middle-dose YRRCS group and high-dose YRRCS group.We made the model of slow transit constipation on all groups but the normal one.According to literature reports,we applied with hypo-dermic injection of hydrochloric acid morphine 2.5mg/kg/d for 45 days.Then the mouse model of constipation was established.Result:The mouse stools of model group turned so sticking and stagnant that it is difficult to fall from anus.That is to say symptoms of mouse of model group is very similar with the patients of constipation.Experiment 2:The influence of YRRCS on defecation function of mouseObjective:To observe the influence of YRRCS on defecation function of mouse,we can judge the efficient effects of YRRCS on slow transit constipation.Method:After establishing the model of slow transit constipation,we began to give the different medicines to the mice of different groups.Normal group and model group were intragastic infused with normal saline.Mazirenwan group was intragastic infused with Mazirenwan.Slight-dose YRRCS group,middle-dose YRRCS group and high-dose YRRCS group were intragastic infused with different dose of YRRCS which contented the medicine of 11.4 g/kg,22.8g/kg and 45.6g/kg.The period of administration was 9 days.Before the ninth day,the mice had been forbided to drink and eat for 12 hours.Then,the mice were intragastic infused as usual.After that,we began to record the quantity and weight of stools in 24 hours.Result:The mouse stools of model group had turned obvious dry and hard.At the same time,the quantity and weight of stools declined evidently,which had significant different with the normal group(P＜0.01).After treating with medicines,all mice stools had recoverd in different level,especially the high-dose group,whose stools had no different with normal group in outward appearance.Conclusions:YRRCS can increase the quanlity and weight of mouse stools of slow transit constipation in 24 hours.The efficient effects depend on the dose of YRRCS.Experiment 3:The influence of YRRCS on the length of ink progradation of the small intestine of mouseObjective:To observe the influence of YRRCS on the length of ink progradation of the small intestine of mouse,we can judge the efficient effects of YRRCS on slow transit constipation.Method:Before the tenth day,the mice of each group had been forbided to drink and eat for 12 hours.Then,all mice were intragastic infused with the medicines which contained the ink. After 30 minites,the mice were killed.Then,we opened the abdomen,cut the small intestine, measured the length of the ink and small intestine.At last,we caculated the rate of ink moving forward.Result:The rate of ink moving forward of small intestine of normal group is 78.20%,while the model group decrease into 65.08%.There are significant different between them(P< 0.05).After treating,the rate of ink moving forward of small intestine of all groups increased especially the high-dose one,who had got to 93.12%.It is not only higher than model group, but also higher than normal group.Conclusions:YRRCS can strengthen the force of stomack and intestine and treat the slow transit constipation.Section two:The Study of Mechanism of YRRCSExperiment 1:The Effects of YRRCS on ICC,SP and VIPObjective:To investigate the mechanism of YRRCS on slow transit constipation treatment by using immunohistochemistry method.Method:After intragastric administration for 2 hours,the mice were anesthetized and excised colon specimen。Then,we used immunohistochemistry method to examine the change of amount and form of ICC,VIP and SP in colon myenteric plexus of slow transit constipation mice model. Result:The amount of ICC,VIP and SP decreased obviously in colon of mouse model with chronic constipation.After treatment by YRRCS,the amount increased more than control group.There are significant different between control group and high-dose group(P＜0.01).Conclusions:YRRCS can strengthen colon peristalsis by increasing the amount of ICC,VIP and SP.So,it can cure slow transit constipation.Experiment 2:The Effects of YRRCS on c-KitObjective:To investigate the mechanism of YRRCS on slow transit constipation treatment by using western blot method.Method:After intragastric administration for 2 hours,the mice were killed and excised colon specimen。Then,we used western blot method to examine the change of amount of c-Kit in colon myenteric plexus of slow transit constipation mice model.Result:Comparing with normal group,the amount of c-Kit in model group decreased obviously(P＜0.01).After treatment by YRRCS,the amount increased 2.95 times than model group,but still lower than normal one(P＜0.05).Conclusions:YRRCS can strengthen colon peristalsis by increasing the amount of c-Kit.So, it can cure slow transit constipation.