| Obgective:the spleen and stomach Damp-heat syndrome is a type of diseases take place in Lingnan,caused by damp-heat,over-eating fatness and pungent,which may injure the spleen and stomach.It is marked with complicated clinical symptoms,longer history and lingering to heal,so in the occurrence and developent of disease,many factors may spur the organism and the body may in turn reflect for the stimulation.This study intend to fabricate the mode of spleen and stomach Damp-heat syndrome in rats by multiple factors including external damp-heat environment,fat and sweet diet and microorganism.We may probe into the behavior change of spleen and stomach damp-heat syndrome,and we also explore the impact on neuroendocrine systyme of hyperactive hypothalamus-pituitary-adrenal axis,the change of adrenal galnd index and the different secretion of CRH,ACTH,COR,β-EP,ALD and ADH.We may probe into mechanism of HPA axis and water-sodium metabolism.In addition,we will get to know the principle of Qingre Qushi of sanren decotion. Methods:1.chinese herbal medicine and foodstuffComposition of sanren decotion:bitter apricot seed,Rhizoma Pinellinae Praeparata,talc,crude semen coicis,ricepaperplant pith,amomum caramomum, bamboo leaf, officinal magnolia bark.High glucose and lipid foodstuff:the ordinary mix with 12% pork lard and 8% honey.2.Groupfifty male Wister rats were randomly divided into five groups:normal control group,model group,sanren decoction high and medium and low dosage groups. Each group has 10 rats.3.creation of modelsThis experiment intend to creat the model of spleen and stomach damp-heat syndrome by compositite factors including external damp-heat environment,fat and sweet diet and microorganism.①Dnorma control group:fed with ordinary foodstuff in 20-28℃and 50-70% relative humid environment,②model group:fed by high suger and fat diet and escherichia coli in simulation of climate warehouse.③feed the rats with low dosage sanren decotion based on the model of spleen and stomach damp-heat syndrome.④feed the rats with medium dosage sanren decotion based on the model of spleen and stomach damp-heat syndrome.⑤fed with high dosage sanren decotion based on the model of spleen and stomach damp-heat syndrome.4.indexs of observation and detectionObserve the change of general state,weight,rectal temperature,diet,drinking and shit character between before and after modeling.Observe the behabior of rats in open-field test by small animal behavior recording and analysis system.Observe respectively the difference of before and after administration.Figure out the index of adrenal gland by weighing method and simultaneously detect the concentration of CRH,ACTH,COR,β-EP,ALD and ADH of rats.5.stastistic methodAll the datas were processed by statistical software SPSS13.0.the data type is quantitative data,expressed by mean±standard deviation(x±s).The data of weight and rectal temperature are analyzed by variance of repeated measure.The data of open-field test is compared by paired t test.The index of adrenal gland and other indexs including CRH,ACTH,COR,β-EP,ALD,ADH are analysed by one-way ANOVA method.Multiple comparison is employed by LSD for different groups, p<0.05 is the significant level.Result:1.general condition of the rats Before the test,all the rats have good condition,active movements,glossy white fur,normal eating and drinking,shape stool and normal temperature.After the test,normal group have not abnormal performance.With the conduct of model establishing,most of rats appear as tired,lazy,dull,fluffy hair,haggard face,reduced diet,sticky stool.From the tenth day,groups employed in composite methods has some abnormal performance,stool is sticky,rectal temperature increases quickly.after entering simulation of climate warehouse,the symptoms are aggravated.the groups with different dose sanren decotion,the movements,diet and stool are improved to norma level.2.the changes of weight and temperatureThe change of weight:there is no statistical significance between every two groups on 1st day(F=0.467,P=0.759).the first ten days,the growth of weight in each group have no difference.Compared with norma group,weight of 4 model groups grow slowly when going into simulation of climate warehouse.The trend of weight gain in model groups keep consistent from 18th to 20th day,but slower than normal group.20 days later,weight growth of Sanren decotion with differtent dose groups return to normal level.It is lower than norma control group at different time point (P=0.000),which prompts weight losss.After giving Sanren decotion,all treatment groups keep grow normaly.Weight of normal control group increase normal all the time.Change of rectal temperature:there is no statistical significance between every two groups on 1st day(F=0.665,P=0.602).the first ten days,the rectal temperature in each group have no significant difference.Retal temperature of 4 model groups when going into simulation of climate warehouse increase quickly.The temperature of model groups go up obviously after giving enteroinvasive E.Coli.Compared with normal group,it has statistic significance(P<0.01).After giving every dosage sanren decotion for 7 days,the rectal temperature of rats decline largely,but model group has no obvious decline.Normal control group has no change at different time point.3.observation of open-field testChange of central cell residence time:befor the treatment,compared with normal group,central cell residence time of model group,sanren decotion high and low dose group prolong significantly(P<0.01),and so sanren decotion does (P<0.05).this reveals that spatial cognitive competenc of rats declines.After the treatment,time of sanren decotion group with different dose is less than that of before the treatment.And compared with model group,it has significant difference(P<0.01 or P<0.05), there is no significant difference between sanren decotion high dose group and medium dose group and low dose group(P>0.05).this shows that sanren decoton can improve the spatial cognitive competence of rats.Change of sports cell and vertical frequency:befor the treatment,compared with normal group,sports cell and vertical frequency of model group,sanren decotion high,medium and low dose group decrease significantly(P<0.01 or P<0.05),this reveals that excitability,capacity to adapt to new environment and competenc to explre of rats declines.After the treatment,sports cell and vertical frequency of sanren decotion group with different dose increase more than that of before the treatment.And compared with model group,it has significant difference(P<0.01 or P<0.05),there is no significant difference between sanren decotion high dose group and medium dose group and low dose group(P>0.05).this shows that sanren decoton can improve the explore behavior and independent activity,and increase excitability of rats.Change of modification frequency:befor the treatment,compared with normal group,modification frequency of model group,sanren decotion high,medium and low dose group decrease significantly(P<0.05),this reveals that awarness and satisfaction to new environment of rats decline.After the treatment,modification frequency of sanren decotion group with different dose increase differently.And compared with model group,it has significant difference(P<0.05),there is no significant difference between sanren decotion high dose group and medium dose group and low dose group(P<0.05),this shows that sanren decoton can improve the degree of satisfaction to new environment of rats.4.detection of experimental indexAfter the success of establishing model,the concentration of plasma CRH,ACTH,serum COR elevates obviously(P<0.01).compared with model group,the concentration of CRH,ACTH and COR in sanren decotion high,medium and low dose group decline obviously and have significant difference(P<0.01 or P<0.05), especially sanren decotion medium dose group has best effect.Sanren decotion medium dose group is better than high dose group to decline CRH,while it is better than low dose group to decline ACTH(P=0.038),while has no difference with high dose group(P=0.091). Medium dose group is better than high dose group to decline Cor(P=0.034),while has no difference with low dose group(P=0.034)Compared with normal group,adrenal gland index and plasmaβ-EP of normal group go up obviously and has significant difference(P<0.01).compared with model group,adrenal gland index and plasmaβ-EP of sanren decotion high,medium and low dose group decreases obviously and has significant difference (P<0.01 or P<0.05).Compared with medium dose group,high and low dose group has significant difference(P<0.01 or P<0.05),but there is no significant difference between high and low dose group(P>0.05).Compared with normal group,ALD and ADH of normal group go up obviously and has significant difference(P<0.01).Compared with model group,ALD and ADH of sanren decotion high,medium and low dose group decreases obviously and has significant difference after giving sanren decotion for 7 days(P<0.01 or P<0.05).Compared with medium dose group,high and low dose group has significant difference to decrease ALD(P<0.05),but there is no significant difference between high and low dose group(P=0.534).there is no significant difference between high,medium and low dose group(P<0.05).Conclusion:The experimental results reveal that under the composite factors of high glucose and lipid foodstuff, damp-heat environment and microorganism,the speen and stomach damp-heat syndrome animal model can be copied successfully and may result in the abnormal change of behavior and secretion of neuroendocrine system.compared with normal control group,the central cell residence time prolong obviously,this reveals that cognitive competence decrease,sports cells,vertical frequency and modification frequency of model group are less than control group,it shows that excitability and the capacity adapt to the new surrounding are lower than control group.And it also reveal that autonomous activities and exploratory behavior of rats in unfamiliar surrounding decrease.Compared with control group, the concentration of plasma CRH,ACTH,secrum Cor in the model rats have a significant difference and lead to the change of the behavior,this show that there are close relation among experimental indexs,behavior and neuroendocrine system,rats are in a stress state,promote to improve self-protection and self-regulation.Growth of adrenal gland index andβ-EP indicates pathological change to adapt to new enviroment in stress state.Several indicators all above may use as uch as indicators of rectal temperature,weight and behavior.The concentration of plasma ADH and secrem ALD increase obviously,reveals that distal renal tubular and collecting ducts reabsorb water enhancely,which may result in moisture stagnation and water-sodium retention.Sanren decotion may have the effects as follows:1)can recover the general condition of spleen and stomach damp-heat syndrome animal model result from composite factors.2) may decrease concentration of ACTH andβ-EP and improve the situation of rats.3)may improve adrenal gland hypertrophy.4)regulate the function of HPA axis and decrease the secretion of CRH,ACTH and Cor.5)regulate secretion of ALD and ADH to maintain constancy of the internal environment.
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