The Study Of Effect And Pharmacology And Investigation Of Effect Of The Placebo Made By Low Concentration Trial Chinese Medicine

Irritable bowel syndrome (IBS) is a function of disorder disease,behaving the abdominal discomfort or pain,being related with abnormal of shape(loose or mucous stool) and change habit of defecation. At present, it is a kind of biologica-psychological-social diseases and relevant with the change of brain-intestine axis (nerve-endocrine-immune network) to change supersensitivity of the relevant organs, gastro-dynamics and the conduct of the mind. Now Western medicine is lack of effective measures of treating this disease. However Chinese medicine tends to achieving good result in treating this disease.Objective:1. The study of exploring the mechanism on diarrhea of IBS rats and pharmacodynamics of Chang an I.2. Observe the Chinese medicine placebo made of 5% and 10% of Chang an I whether the pharmacologic effect from gastrointestinal motor,analgesia and anti-inflammatory experiments and investivigation of pharmacological effect on D-IBS rats.Method:First:The study of normal pharmacological effect on D-IBS by Chang an I.Methods:To observe the Pharmacodynamic effects of Chang an I by the experiments of intestinal propel and accentuation by neostigmin,analgesia and anti-inflammatory experiments.Result:The high,medium and low dose of Chang an I had statistical significance compared with normal group (P＜0.05) in the experiments of intestinal propel and accentuation by neostigmin and anti-inflammatory experiments, and there were no differences among the three groups (P>0.05);The analgesia of hot plate,the high,medium group could improve the pain threshold when 30',60 'and 90',120' after medicine and there were no different each other (P> 0.05),the change of pain threshold had no statistical significance compared with normal group (P> 0.05) when 30'after medicine.Second:the study of mechanism on D-IBS rats by Chang an I.method:Wistar rats were ramdomly divided into six groups:normal group,model group,high,medium and low dose group and Pinaverium Bromide group, each group of 10. Fasted, free drunk water 10 hours before experiment,senna leaf (0.3g/mL) i.g(20ml/kg),qd.After 1 hour ig bundled the rat two limbs with the cotton bondage. Which caused a certain stimulate to make them dysphoria,Lasted two hours,for two weeks.Given medicine after model achieved, normal group only 0.5%(CMC) mixture of the fluid (20ml/kg).Given medicine 60'after senna leaf i.g 2weeks later:the dose of high,medium,low of Chang an I:22.6g/kg,11.3g/kg,5.7g/kg, Pinaverium Bromide group with Pinaverium Bromide, the volume of medicine was 20ml/kg,qd,last 2 weeks.To study the mechanism of Chang an I on D-IBS rat from the rat high sensitivity,diarrhea index, an inflammatory media,the immune functions. pathological changes and protein expression.Experiment one:The influence of Chang an I on visceral high sensitivity of D-IBS rat.Observe the influence of Chang an I on visceral high sensitivity of D-IBS rat with the AWR scores measured pain reaction under CRD of 20,40,60,80mmHg.Experiment two:The influence of Chang an I on serum cytokine of D-IBS rat.With typical filling enzyme immune sorption measure, the steps of measure of serum IL-10,TNF-a strictly followed the request of Kit.Experiment three:The influence of Chang an I on immune function of D-IBS rat.After experiment,the rats fasted 24h and weighed and calculated the thymus and spleen index.Experiment Four:The influence of Chang an I on Mast Cell in colonic mucosa of D-IBS ratObservers the number, area, the circumference and diameter. of MC by ALLen neutral red staining technique.Experiment Five:The influence of Chang an I on NF-κB and it's protein expression.The method of immunohistochemistry and westernbloting print measure the content of NF-κB in colonic mucosa of D-IBS rat and it's protein expression.Result(1) the establishment of D-IBS rat model1) The changes of weight and appearance.:there were loose stool,slack coatfur and feather, myasthenia of limbs,taken in sail,thin,etc. The weight of animals model group increased slowly compared with normal group were statistical significant difference after molding 1 and 2 weeks (p<0.05).2) The first day after animal model establishment, diarrhea index of trial groups compared with normal group had statistical significance (P＜0.01) and han no statistical significance among them,prompted the model established; The reduce of diarrhea index had statistical significance compared with model group (P＜0.05).3) The AWR score of trial group had statistical significance compared with normal group under different pressure with CRD after model establishment (P＜0.01-0.05).4) The result of HE dyschroia in rat colonic mucosa. Colonic mucosa were intact,There were only two rats minor vasodilation,no physical lesion and inflammatory cells infiltration obviously..5) The thymus and spleen index of model group all decreased compared with normal group.6) The reduce of serum IL-10 content and increase of serum TNF-a content of model group compared with normal group.(2) The influence of visceral high sensitivity of D-IBS rat.The AWR score of Chang an I medium,low dose had no statistical significant compared with model group(P>0.05), high dose of Chang an I compared with model group(P<0.01),The high,medium,low dose of Chang an I had statistical significant compared with model group (P＜0.01-0.05), been dose-response relationship.(3) he influence of Chang an I on serum cytokine of D-IBS rat.At the end of treatment,the serum TNF-a down with the content of high dose of Chang an I had statistical difference compared with model group (p< 0.01), medium and low dose down with the content of group model of a differential (p<0.05); the serum IL-10 up with the content of high. medium,low dose of Chang an I had statistical difference compared with model group (p <0.05),Treatment at the end of the high doses, and i set down, and content of a model of a differential (p<0.01), medium and low dose down with the content of group model of a differential (p< 0.05); model set the il-10 level below the normal group (p< 0.05), the end of the intestine anni, high, medium and low dose of il-10 increase in the model of a differential (p<0.05); serum il-10 content with a model of any differences (p.0.05). the number and i is high, medium and low(4) he influence of Chang an I on immune function of D-IBS rat.1) Weight:The high,medium,low dose of Chang an I could obviously improve the weight of rat when 1.2weeks after treatment, there were statistical significance((P＜0.05 or P＜0.01).2)Thymus. spleen:Increasement of index of thymus,spleen on high,medium,low dose of Chang an I when treatment ended,there were statistical significance compared with model group (P<0.05).There were no statistical significance among these groups.(5) The influence of Chang an I on NF-κB and it's protein expression.1)The result of immunohistochemistry:The content of NF-κBP65 in normal group were no or weak positive expression, the color were brown,mainly in the cytoplast. Chang an I reduced the expression compared with model group been statistical significant compared wtith model group (P＜0.05),mainly in the cytoplast. There were no statistical significant among these groups.2) The result of Westernblot:Chang an I reduced the NF-κBP65 protein expression compared with model group been statistical significant compared wtith model group (P＜0.05).There were no statistical significant among these groups.Third:The study of the normal pharmacodynatics placebo made from low concentration of trial medicine for IBSOne:The study of normal pharmacological effect on D-IBS by the placebo.Methods:To observe the Pharmacodynamic effects of the placebo by the experiments of intestinal propel and accentuation by neostigmin,analgesia and anti-inflammatory experiments.Result:The placebo of 5%,10% Chang an I had no statistical significance compared with normal group (P> 0.05) in the experiments of intestinal propel and accentuation by neostigmin. analgesia of hot plate and anti-inflammatory experiments.Two:the study of mechanism on D-IBS rats by placebo.Method:Experiment one The influence of the placebo on visceral high sensitivity of D-IBS rat.Observed The influence of the placebo on visceral high sensitivity of D-IBS rat with the AWR scores measured pain reaction under CRD of 20,40,60,80mmHg.Experiment two The influence of the placebo on serum cytokine of D-IBS rat.With typical filling enzyme immune sorption measure, the steps of measure of serum IL-10,TNF-αstrictly followed the request of Kit.Experiment three:The influence of the placebo on immune function of D-IBS rat.After experiment,the rats fasted 24h and weighed and calculated the thymus and spleen index.Experiment Four:The influence of the placebo on Mast Cell in colonic mucosa of D-IBS ratObserved the number,area, the circumference and diameter. of MC by ALLen neutral red staining technique.Experiment Five:The influence of the placebo on NF-κB and it's protein expression.The method of immunohistochemistry and westernblotirig print measure the content of NF-κB in colonic mucosa of D-IBS rat and it's protein expression. Result:1)The influence of visceral high sensitivity of D-IBS rat.The AWR score of the placebo of 5%,10% Chang an I had no statistical significant compared with model group(P>0.05).2) The influence of Chang an I on serum cytokine of D-IBS rat.At the end of treatment,the serum TNF-a down with the content of the placebo of 5%,10% Chang an I had no statistical significant compared with model group(P>0.05).the serum IL-10 up with the content of the placebo of 5%.10% Chang an I had no statistical significant compared with model group(P>0.05).3) the influence of Chang an I on immune function of D-IBS rat.The increase of thymus and spleen index of placebo of 5%.10% Chang an I had no statistical significant compared with model group(P>0.05).4) The influence of Chang an I on NF-κB and it's protein expression.1)The result of immunohistochemistry:NF-κBP65in the placebo of 5%,10% Chang an I had no statistical significant compared with model group(P>0.05). NF-κBP65 of two groups expressed mainly in the nucleus.2) The result of Westernblot:Chang an I reduced the NF-κBP65 protein expression o f the placebo of 5%,10% Chang an I had no statistical significant compared with model group(P>0.05). ConclusionsChang An I, high, medium and low-dose group and its low concentration of the original drug to placebo in the gastrointestinal motility, analgesic and anti-inflammatory experiments, as well as in research on visceral sensitivity in IBS-D rats and diarrhea index, inflammatory cytokines TNF-a, IL-10 and colon by immunohistochemistry the expression of NF-κB and its impact results showed that intestinal-an I, high, medium and low-dose group, pharmacodynamic effects and indicators for each group from the above Compared with model group, results showed that the above indicators can be described by an I, treatment of irritable IBS-D rats were part of the mechanism.5%,10% none of the above pharmacodynamic effect on the IBS-D in rats has no effect on the above indicators of the mechanism can play the role of traditional Chinese medicine, placebo-controlled.An article on the intestinal first-I and its low concentration of the original drug to placebo in efficacy studies as well as IBS-D model of part of the mechanism of rat studies,using multi-factor (senna combined tradition-bound method) working together, from the perspective of different factors together induce IBS-D generation, mainly through low-grade inflammation in IBS-D pathological characteristics of study commence in low-grade inflammation in the existence of inflammatory bowel cytokines, and NF-κB activation, causing visceral sensitivity, abnormal gastrointestinal motility, and thus to explain Chang An I, on the part of the IBS-D rats pathogenesis.