| Objective By observing the changes of "AVP-V2R-cAMP-PKA-AQP2 pathway" and the effects of Bixiefenqingyin(abbreviation:BX)on water metabolism disorder in kidney-yang deficiency rats after given the V2R inhibitor OPC-31260 and PKA inhibitor H-89,to reveal the mechanism of metabolism disorder in kidney-yang deficiency rats and the role targets of BX.Methods(1)The kidney-yang deficiency rats(n=70)were treated with adenine(200mg/kg)once per day for 21 days,while the normal group(n=10)given the same volume of saline.The anal temperature,holding power,weight and general symptoms and signs of kidney-yang deficiency including erect hair,back arched,curled up,grab irritation were observed per 4 days,drinking,urine and diet were detected per week.The urine of third week was collected to detect the content of 24 hour U-TP and 17-OHCS.Serum was centrifuged by Orbital blood to detect the content of ACTH,CORT,BUN,Scr.Compare the all indexes to decide whether the model was successful.(2)Then regroup the model group into the kidney-yang deficiency model group,model and V2R inhibiting group,model and PKA inhibiting group,BX group,BX and V2R inhibiting group,BX and PKA inhibiting group.All the BX groups were treated with 24.3 g/kg raw herbs of BX soup per day for 21 days,The V2R inhibiting groups were treated intravenously with OPC-312605mg/kg per day for the last 3 days,the PKA inhibiting groups were treated intravenously with H-89 400ug/kg per day for the last 3 days.(3)The anal temperature,holding power,weight and general symptoms and signs of kidney-yang deficiency including erect hair,back arched,curled up,grab irritation were observed per 4 days,drinking,urine and diet were detected per week and the 4 hours urine was also detected in the last week.The urine of third week were collected to detect the content of 24 hour U-TP and 17-OHCS.On the 21th,rats were anesthetized,and blood was collected by heart punctures and separated for serum to detect the contents of ACTH,CORT,AVP;hypothalamus homogenates were prepared for the detection of the content of CRH;renal medullar homogenates were prepared for the contents of V2R,cAMP,PKA,AQP2,Epac,Ca2+.All the detections in homogenates were performed by ELISA assay.Results(1)Compared with the normal contral group,the kidney yang deficiency model rats have the following symptoms:the weight grew slower,anal temperature decreased,holding power decreased,urine increased and general symptoms for signs of kidney-yang deficiency including erect hair,back arched,curled up,grab irritation came out in the model group,and the content of ACTH,CORT in the serum and 17-OHCS significantly decreased,the content of BUN,Scr,U-TP significantly increased.(2)After the treatment of BX soup for 21 days,the syndrome of kidney-yang deficiency improved such as weight grow faster,anal temperature,holding power higher and so on.The content of ACTH,CORT,17-OHCS,V2R,cAMP,PKA,AQP2 all significantly increased and the content of Scr,U-TP all significantly decreased compared with the model group.(3)Treatment with V2R inhibitor and PKA inhibitor intravenously for the last three days,the content of AVP,V2R,cAMP,PKA,AQP2 were significantly decreased.(4)Treatment with V2R inhibitor and PKA inhibitor intravenously for three days,there were no significant difference in the content of A VP,V2R,cAMP,AQP2,PKA between the BX soup groups.Conclusion(1)By the comprehensive comparison of various evaluation indicators,it is found that the method of adenine administered intragastrically for 21ds(200mg/kg/d)can successfully build the kidney-yang deficiency model.(2)BX soup can effectively improve the general symptoms and signs of kidney-yang deficiency rats,can up-regulate the content of AQP2 and correct the water metabolism disorder of kidney-yang deficiency.(3)The effect of BX soup correcting the water metabolism disorder of kidney-yang deficiency is related with the multi-targets regulating "AVP-V2R-cAMP-PKA-AQP2 pathway". |
PDF Full Text Request