The Experimental Study On Quantizing Dominant Wet And Dominant Heat Type Of The Spleen And Stomach Wet Heat Syndrome

Objective:The Spleen and Stomach Wet Heat Syndrome(SSWHS) is the frequent syndrome, it's hard to diagnose,because its pathological change includes dominant wet(DW) syndrome,dominant heat(DH) syndrome and damp-heat syndrome.To study the essence of SSWHS,SD rat was taken as researching object.We research the rat models of DW syndrome and DH syndrome by means of many factors including high glucose and lipid feeder,wet and heat environment,pathogen and so on.We also researched the mechanism of action of Sanren Tang(ST) and Baihucangzhu Tang(BT) on DW syndrome and DH syndrome rat models.General condition and a series of experimental index change of rat were observed.According to the results,we approached the essence of DW syndrome and DH syndrome,the mechanism of action of ST and BT.The study result provided experimental evidence for quantization of DW syndrome and DH syndrome. Methods:1.Chinese herbal medicine and feederComposition of ST:amygdale amara,prepared rhizome pinelliae with aulmen radix glyrrhigae calcaren,musslinite,coicis semen,amomi cardamomi,magnolia bark,medulla terapanacis,lophatheri.Composition of BT:cypsum fibrosum,anemarrhenae,rice fruit,glycyrrhizae.Composition of Fuziganjiang Tang(FT):aconite root,zingiberis.High glucose and lipid feeder:the ordinary feeder mix with 12%hog fat and 8% honey.2.Group40 SPF grade SD rats,including 20 female and 20 male,were divided into five groups(each group 8 rats) randomly,the normal control group(NCG),DW group (DWG) and ST group(STG),DH group(DHG) and BT group(BTG).3.Creation of modelsThe DW syndrome and DH syndrome of the SSWHS rat models set up by the composite factors including high glucose and lipid feeder,wet and heat environment, pathogen and and so on.①NCG:fed by ordinary fodder in 20-28℃temperature and 50%-70%relative humidity environment;②DWG:high glucose and high lipid feeder +climate box + wine + colibacillus;③STG:high glucose and high lipid feeder +climate box + wine + colibacillus +ST;④DHG:high glucose and high lipid feeder +climate box + FT + colibacillus;⑤BTG:high glucose and high lipid feeder +climate box + FT + colibacillus +BT.4.Indexs of observation and detectionBefore and after the creation of models,changes of general condition including weight,temperature,24 hours urine volume and sedes description were contrasted. We detected aquaporin2(AQP2) in urine with the method of ELISA,(Heat Shock Protein70(HSP70) in homeocyte of blood plasm with flow cytometer,the content of Substance P(SP),Somatostatin(SS) sinus ventriculi with radioimmunoassay. Executed the rats,took out of gastro and duodenum tissue.Then the tissue were put into 10%formalin,imbedded by paraffin and made blade,stained tissue were put into 10%formalin,imbedded by paraffin and made blade,stained with HE and observed under light microscope.5.Statistics methodAll the data were processed by statistical software SPSS13.0.The data type was quantity data,expressed by mean±standard deviation((?)±s).The data of weight, temperature were analyzed by variance of repeated measure.The data of 24 hours urine volume,the content of AQP2,HSP70,SP and SS were analyzed by One-Way ANOVA.Multiple comparison was by LSD for different groups.P＜0.05 is the significant level.Result:1.General condition of the ratsNCG:During the period of four weeks,the rats had luster hair,good appetite, normal activity and sedes.DWG and DHG:As time was going on,most of rats were tired,enjoyed to lie down,acted lazily,reacted slowly,and their hair was fluffy and withered.From the seventh day,all rats had loose stool or sticking and greasy sedes. After moved into the climate box,the symptom,aggravated.The rats had much severe loose stool or sticking and greasy sedes,and their crissum filth appeared. These appearance of DWG was most obviously.STG and BTG:The rats appeared similar symptom as DWG and DHG,but the symptom relieved after drenched by ST and BT.2.The changes of weight,temperature and 24 hours urine volume The weight of the 4 model groups were decreased after moved into the climate box.Their weight were lighter than the NCG at different point of time(P＜0.01), while the weight of the STG and BTG can recover to the normal level after treated by ST and BT.The temperature of model groups stepped up after they were put into the climate box,and stepped up much rapidly after drenched the colibacillus.The result showed significant difference when compared with the NCG(P＜0.05).The temperature of the STG and BTG stepped down to the normal level after treated by ST and BT,while the temperature of the DWG and DHG never stepped down.The 24 hours urine volume of the DWG and DHG decreased.The result showed highly significant difference when compared with the NCG(P＜0.01).There was no significant difference about the 24 hours urine volume when the STG and BTG compared with the NCG(P＞0.05).3.Result of experimental indexsThe content of AQP2 of the DWG and DHG were significantly higher than the NCG(P＜0.01),while the content of the DWG was much higher than the DHG (P＜0.01).The content of AQP2 of the STG and BTG can recover to the normal level, the result showed no significant difference when compared with the NCG(P＞0.05).The expression of HSP70 in homeocyte of blood plasm of the DWG and DHG were higher than the NCG(P＜0.001),while the expression of the DHG was much higher than DWG(P＜0.01).The expression of HSP70 of the STG and BTG can recover to the normal level,the result showed no significantly difference when compared with the NCG(P＞0.05).The content of SP,SS in sinus ventriculi of the DWG and DHG decreased when compared with the NCG(P＜0.001).The contrast of SP,SS in DWG was much lower than DHG(P＜0.01).The content of SP,SS of the STG and BTG can recover to the normal level,the result showed no significant difference when compared with the NCG(P＞0.05).4.Pathological section of Gastro and duodenum tissue In the DCG,there was little inflammatory cell at mucous layer of gastro,the endepidermis of the intestinal villus of duodenum was integrity.There was no obviously significant difference between the DWG and DHG about the pathological section of gastro and duodenum tissue,which showed:(1) at mucous layer of gastro,a great quantity of heterophil granulocyte,eosinophile granulocyte,anapetia and congestion can been seen;the submucous layer of stomach was dropsical,heterophil granulocyte and eosinophile granulocyte also can been seen.(2) the cellula epithelialis of the intestinal villi of duodenum was deciduous,a great quantity of inflammatory cell also been seen.From the section of the STG and BTG,we can see less inflammatory cell,no anapetia and congestion,no dropsy in the submucous layer of stomach,regular endepidermis of the intestinal viili of duodenum,and just a little inflammatory cell.Conclusion:The experimental result reveal that under the composite factors of high glucose and lipid feeder,wine,wet and heat environment,pathogen and so on,the DW syndrome and DH syndrome animal models can be copied successfully.The content of AQP2 of the DWG and DHG increased,and the DWG increased much higher than the DHG.This result revealed that the physiological function of evacuating water was abnormal,so AQP2 is an important index to express how the "wetness" from and quantize wetness.The expression of HSP70 in homeocyte of blood plasm of the DWG and DHG increased,and the DHG increased much than the DWG.The result revealed that the rats were in the condition of self- protection and self-adjustment under the composite factors.HSP70 can be taken as one of the important index to quantize DW syndrome and DH syndrome.The content of SP,SS in sinus ventriculi of the DWG and DHG decreased.The contrast of SP,SS in DWG was much lower than DHG.The content of SP,SS can explain the clinical symptom of digestive system about s DW syndrome and DH syndrome.The gastro and duodenum of the DWG and DHG occur inflammatory pathological changes,revealing that the clinical symptom of the spleen and stomach wet heat syndrome had its own structural foundation.ST and BTmay have the effect as follow:1) can recover the sings and DW syndrome and DH syndrome animal models which was caused by the composite factors,2) regulate the content of AQP2,recover the normal physiological function of evacuating water,3) decrease the increased expression of HSP70 caused by heat stress, 4) regulate the content of SP and SS in sinus ventriculi,recover the normal stomach intestine function,5) partly repair the structure of gastro and duodenum of rat models.