Effects And Mechanism Of Danggui-shaoyao-san On Nephrotic Syndrome Based On The Path Of AVP-V2R-AQP2

Aim:To research the pharmacological treatment and mechanism of Danggui-Shaoyao-San(DSS) on nephritic syndrome. To evaluate the function of DSS on rats model with nephritic syndrome, in the 24 h urine protein, renal function, renal tissue pathology morphology and electrolyte metabolism as incaditor. We discuss the mechanism of DSS on nephritic syndrome, through the pathways AVP-V2R-AQP2 on the basis of preliminary studies. The research can provide the experiment basis and function of DSS applied in clinical treatment on nephritic syndrome.Methods:(1)90 male SD rats were devided into normal group,model group,the high dose、medium dose、low dose of Danggui-Shaoyao-San group,Fosinopril group; The methods of adriamycin solution injected to the caudal vein of rats is adopted to establish the rat model with nephritic syndrome except normal group. After the successful modeling,the each group of Danggui-Shaoyao-San were fed with Danggui-Shaoyao-San high、medium and low dose and the Fosinopril group were fed with fcinepoils for 4weeks.(2)Metabolic cages were used to collect 24 hours urine of rats, 24 h urine protein content was measured by BCA method.(3)The levels of serum BUN、Cr、TC、TG、TP、ALB were detected by the automatic biochemistry analyzer.(4)The level of Na+、K+ 、 Cl- in serum and urine were detected by the automatic biochemistry analyzer.(5)The glomerular and renal tubular pathological changes were observed by HE staining in renal tissue.(6)The level of ET-1、Ang II、AVP in plasma were detected by enzyme-linked immunoabsorbent assay(ELISA).(7)The relative expression of AQP2 gene and V2 R gene were determined by RT-PCR.(8)The relative expression of AQP2、(AVP)V2R protein in renal tissue were detected by Western Blotting.(9)The levels of AQP2 in urine were detected by enzyme-linked immunoabsorbent assay(ELISA).Results:(1)After the model injection of doxorubicin 0.6mg / 100 g in tail vein 1、2 weekslater. The 24 h urine in rats were significantly decreased,the 24 h protein in urine were increased significantly, the renal tissue disease occurs,the model of Nephrotic Syndrome were build success.(2)Compared with model group, the level of 24 h urine protein were decreased significantly in treatment group.(3)The renal function level of BUN, Cr in serum have different degrees of decline, TC、TG in serum have different degrees of decline. TP, ALB in serum decreased significantly.(4)The level of Na+、K+、Cl- in serum increased greatly, while level of Na+ 、 K+ 、 Cl- in urine were decreased significantly.(5)Compared with model rats, the degree of pathological changes in glomerular, tubular lesions have improved significantly in treatment group.(6)The level of AQP2 in urine have increased greatly.(7)The level of ET-1、Ang II、AVP in plasma have reduced significantly in treatment group.(8) The expression of AQP2-m RNA 、(AVP)V2R-m RNA reduced clearly, meanwhile, the expression of protein AQP2 and(AVP)V2R descend in treatment group.Conclusions: The stable model of rats with nephritic syndrome can reproduced by the methods of adriamycin solution injected to the caudal vein. Danggui-Shaoyao-San have treatment on rats with nephritic syndrome include improving renal function, reduced the24 h urine protein, and have a certain corrective action on electrolyte disorders, and delaying nephrotic syndrome pathological process. Its mechanism may be related to arginine vasopressin system and AQP2.