Pharmacodynamics And Mechanism Of Action Research Based On The Processing Principle Of Bran Fried Rhizoma Atractylodis Slices

Objective The research on the basis of previous studies,to further explore the differences between before and after pharmacological action of bran fried Atractylodes by using modern analytical techniques,and further processing technology of Rhizoma Atractylodis,lay a foundation for establishing the quality standard of Rhizoma Atractylodis,through pharmacological experiments,the effects of processing on Atractylodes efficacy,changes of the chemical composition and the relationship between system.Methods Chapter One : Comparative study of pharmacodynamics of rhizoma atractylodis before and after processing:60 Wistar rats,Evenly divided male and female,were randomly divided into normal group,model group,rhizoma atractylodis raw product high and low dose group,bran fried rhizoma atractylodis high,low dose group.The preparation of rhizoma atractylodis extracts.Wet resistance medium syndrome animal model is set up.After last time lavage dosing,measure rate of small intestine propulsion and stomach retention rate.Phloroglucinol method for serum,urine,D-xylose content of rat,ria method detecting gastric secrete element(GAS)in the serum,vasopressin(AVP),vasoactive intestinal peptide(VIP),urine water channels protein 2(AQP2),ELISA method to detect the contents of each part of the gastrointestinal tract of water channel proteins.The second part:The effect of bran Fried rhizoma atractylodis on wet resistance medium rats the MAPK signaling pathways and colon AQP3,AQP4 expression.Chapter Tow : The First section :The effect of bran Fried rhizoma atractylodis extract on MAPK pathway of wet resistance medium rats colonic tissue:Building after the immunohistochemical method for wet resistance medium ERK in rat colon MAPK pathway,p38 lightning MAPK protein expression were determined.Rt-pcr method to determine the wet resistance medium ERK in rat colon MAPK pathway,p38 lightning MAPK m RNA expression.The Second Section: After the MAPK signal access is blocked,the effect of bran Fried rhizoma atractylodis on wet resistance medium rat colon tissue AQP3 and AQP4 expression:Building after the Western blot method for wet resistance medium rats colon AQP3,AQP4 protein expression were determined.Rt-pcr method wet resistance di rats colon AQP3 and expression of AQP4 m RNA.Chapter Three :The processing mechanism analysis and research Bran Fried rhizoma atractylodis Indicator component.The First section :pharmacology study of rhizoma atractylodis :Ria method were determined for rat serum IL-1,2,6,and the content of TNF alpha.Rt-PCR method determine the stomach tissue MTLR m RNA,jejunum VIPR2 m RNA expression and quantitative analysis,and detect the influence atractylodes rhizoma atractylodis in bran Fried rhizoma atractylodis on colonic mucosa tissue pathology and the intestinal mucosa tissue ultrastructure of wet resistance medium rat model;The Second section :harmacological research of 5-HMF :Radioimmunity method was used to determine the effect of 5-HMF in bran Fried rhizoma atractylodis on the serum MTL,SP and SS content of wet resistance medium model rat,colorimetric method determined the influence of 5-HMF in bran Fried rhizoma atractylodis on jejunum group ATP content,Immunohistochemical method determined smooth muscle cells in gastric antrum protein epidermal growth factor receptor(EGFR)and proliferating cell nucleus antigen(PCNA),TFF1 in gastric mucosa tissue and the amount of kit expression in the colon tissue of model rats.Results The results of The first part: Compared with model group,stomach retention rate of decreased obviously,small intestine propulsion than increased significantly in the extracts high and low dose group from rhizoma atractylodis and bran Fried rhizoma atractylodis(P < 0.05 or P < 0.01),compared with model group,rhizoma atractylodis extract of rhizoma atractylodis and bran fry rhizoma atractylodis high and low dose group serum,urine D-xylose content significantly increased,the content of AQP2 significantly decreased(P < 0.05 or P < 0.01),rhizoma atractylodis and bran Fried rhizoma atractylodis extract high and low dose group of rats serum AVP,GAS content higher in different degree,VIP content in the different degree lower(P < 0.05 or P < 0.01),compared with model group,rhizoma atractylodis and bran Fried rhizoma atractylodis extract high,low dose group stomach and intestinal AQP2 content significantly decreased(P < 0.05 or P < 0.01).The results of the second part :The results of experimental one :(1)Compared with model group,and bran Fried rhizoma atractylodis extract group of colon tissue of ERK expression significantly reduced(P < 0.01),the group of colon tissue p38 MAPK bran Fried rhizoma atractylodis extract protein expression decreased significantly(P < 0.01),bran Fried rhizoma atractylodis extract group rats colon tissue ERK m RNA expression reduced significantly(P < 0.01),bran Fried rhizoma atractylodis extract group of colon tissue p38 MAPK m RNA expression significantly reduced(P < 0.01);The results of experimental two :(1)Compared with bran Fried rhizoma atractylodis extract group,U0126 + bran Fried rhizoma atractylodis extract group and SB203580 + bran Fried rhizoma atractylodis extract group rats colon tissue AQP3,AQP4 protein expression significantly decreased(P < 0.05 or P < 0.01),Compared with bran Fried rhizoma atractylodis extract group,U0126 + bran Fried rhizoma atractylodis extract group and SB203580 + bran Fried rhizoma atractylodis extract group rats colon tissue AQP3,AQP4 m RNA expression significantly decreased(P < 0.05 or P < 0.01).The results of the third part :The results of experimental one :(1)Compared with model group,in rhizoma atractylodis,high,medium and low dose group domperidone group rats the serum cytokine IL-1,2,IL-6,IL-TNF alpha express different level increases(P < 0.05 or P < 0.01)(2)Compared with model group,rhizoma atractylodis high,medium and low dose group,domperidone group rats stomach tissue MTLR m RNA expression of relative quantity are different degree rise,significant differences(P < 0.05 or P < 0.01),rhizoma atractylodis high,medium and low dose group,domperidone group rat jejunum tissues VIPR2 m RNA expression of relative quantity is reduced in different degree,significant differences(P < 0.05 or P < 0.01);(3)Model group rats intestinal mucosa and submucosa hemorrhage,edema,big flake off,rhizoma atractylodis high dose group rat intestinal mucosa and submucosa hyperemia edema is light.Model group rats colon mucosa epithelial cells volume increase,swelling,membrane of certain enzyme activity change,rhizoma atractylodis high dose group of rats colon mucosa epithelial cells volume has returned to normal.The results of experiment two :compared with model group,5-HMF high,medium and low dose group,domperidone group rats serum MTL,SP and SS content different degree increased(P < 0.05 or P < 0.01),in 5-HMF high,medium and low dose group,the domperidone group ATP express different were degree increased in rat jejunum epithelial cells(P < 0.05 or P < 0.01),5-HMF high,medium and low dose group,domperidone group rats gastric antrum smooth muscle cells in the amount of the expression of EGFR were different degree increased(P < 0.05 or P < 0.01),5-HMF high,medium and low dose group,domperidone group rats gastric antrum smooth muscle cells in the amount of PCNA expression were in different degree increased(P < 0.05 or P < 0.01),5-HMF high,medium and low dose group,domperidone group rat gastric mucosa gland cell cytoplasm in the expression of TFF1 were different degree increases(P < 0.05 or P < 0.01),5-HMF high,medium and low dose group,domperidone group rats colon tissue c-kit have a different degree increases expression(P < 0.05 or P < 0.01);Conclusion:(1)Comparative study of pharmacodynamics before and after the processing of Atractylodes Astragalus gluten can significantly reduce the intragastric residue rate and significantly increase the small intestine propulsion rate in model rats,increase the AVP and GAS content in model rats,reduce the content of VIP,and reduce the expression of ERKm RNA in rats.The result suggests that Astragalus gluten After stir-frying,the spleen function is enhanced.In addition,the content of AQP2 in the urine of model rats was significantly reduced compared with that of gluten,suggesting that the desiccation and dampness of asparagus were eased after stir-fry.(2)Effect of Gluten Astragalus Extract on MAPK Signal Path and Expression of AQP3 and AQP4 in Colon of Wet Resistance Medium Coked Rats.AQP3 and AQP4 expression of colon in wet resistance medium focal rats could be significantly enhanced by Gluten Sophora extract.After the intervention of SB203580 and U0126,the gene and protein expression of AQP3 and AQP4 were significantly decreased in the colonic rats.(3)Study on the Mechanism of Key Variable Components of Gluten Fried AgaricusThe expression of ascosin in the jejunum VIPR2 gene and the expression of MTLR gene in the gastric sinus were significantly increased in rats with wet resistance medium focal syndrome model,indicating that ascorbic can obviously correct the abnormal levels of the VIP2 and MTLR genes.The results of pathological tests can be seen that,Atractyl also has a good recovery effect on the organic damage of gastrointestinal tract in rats with wet resistance medium focal model.This effect may be achieved through the comprehensive effect on gastrointestinal hormones and immune functions of model rats.It is proved that Atractyl is one of the components of Atractyl’s spleen.5-HMF can improve the expression of gastrointestinal hormones and related cytokines in model rats,and improve the gastrointestinal tissue structure of rats from tissue to cell level,indicating that 5-HMF is also one of the important components of strengthening spleen function after asparagus bran stir-fry.