| Object:To explore the effects of Li-zhong decoction on small intestinal peptide transporter(PepT1) in Spleen-Yang deficiency rats,and regulate factors connecting them.Methods:1) Set up the spleen-yang deficiency rat model:Using the composite factors method to set up the spleen-yang deficiency rat model for 3 weeks on male SD rats,which includes intragastric administration of cold senna water at 10g/kg everyday,hypodermic injection apoplon at 0.5mg/kg every other day,and swimming for lOmins every the other day. Verifying models based on manifestation,urine D-xylose excretory rate, andultramicrostructureunder electron microscope.2) Observe the curative function of Li-zhong decotction:Male SD rats were deivded into 2 groups: model group and blank group.The model group rats were bearing the composite factors method to set up the spleen-yang deficiency rat model,and the blank group rats were giving subcutaneous injection and intragastric administration with saline but without swimming for 3 weeks.Stopping all of the factors and giving part of the model rats Li-zhong decoction and the rest water as well as the blank group rats for 4 weeks.Evaluating function of Li-zhong decoction based on manifestation,urine D-xylose excretory rate,and ultramicrostructure under electron microscope.3) Set up spleen yang deficiency rat models with the composite factors method and select parts of normal rats and model rats to observe translating activity of PepT1 by RP-HPLC detecting the blood concentration of cefalexin,which has the similar chemical construction with peptide,after being administrated in duodenum with pylorus ligated.Detect PepT1 protein in small intestine mucosa by way of Western Blot.Detect PepT1 mRNA by way of RT-PCR.Detect influential factors of PepT1 such as cAMP,TNF-α,IFN-γand leptin by way of ELISA.Give model rats with or without Li-Zhong decoction for 4 weeks and observe the same projects.According to different experiment purposes,serum and mucosa of small intestine were collected.Results:1) Spleen-yang deficiency rats modeled with the composite factors method for 3weeks were thin and lassitude with poor appetite,dirty fur,loose stool,low temperature,decreased urinary D-xylose excretory rate,and construction changes of chondrosome with swell,collapse or disappear of cristae.2) The blood concentration of cefalexin was higher of spleen-yang deficiency group than that of normals(P＜0.01),which had no difference with normal group and groups treated with or without Li-Zhong decoction(P＞0.05).3) Protein of PepT1 in spleen-yang deficiency group higher than the normal group(P＜0.01),and that of the group treated with Li-Zhong decoction recovered to have no difference with the normal(P＞0.05), which still remain higher than the normal that of the group treated without Li-Zhong decoction(P＜0.05).4) There was no difference of mRNA between groups normal and spleen-yang deficiency,as well as between groups treated with or without Li-Zhong decoction(P＞0.05).5) cAMP in intestine mucosa of spleen-yang deficiency was higher than that of normal(P＜0.05),which was much lower in rats treated without Li-Zhong decoction than that of rats treated with decoction(P＜0.05).Conclutions:1) The composite factors method modeling spleen yang deficiency rats is suitable for pharmacology study of Li-zhong decoction evaluated by manifestation,urinary D-xylose excretory rate, ultramicrostructure of cellula columnoepithelialis.2) Li-zhong decoction has the function of improving absorption of small intestine which was verified by relieving manifestation of spleen yang deficiency,increasing urinary D-xylose excretory rate,ameliorating ultramicrostructure of cellula columnoepithelialis in small intestine.3) Li-zhong decoction treating malfunction of absorption of small intestine in spleen yang deficiency may through way of cAMP-PepT1 regulatory road. |
