Study On Activating Blood And Promting Water Effects Of Danggui Shaoyao San On Rats With Cirrhotic Ascites

Objective: To investigate the pharmacological effects and mechanism ofDanggui-Shaoyao-San （DSS） on liver cirrhosis ascites, which is the classic symptomsof unsmmoth blood circulation results in water retention. To evaluate the function ofDSS and its decompositions on rats mode with liver cirrhosis ascites, separately fromthe liver and kidney function, liver fibrosis indexes and tissue pathological morphologyand electrolyte metabolism. Examining the effects of the activating blood component（which compsed of angelica, peony, chuanxiong） and the promoting watercomponent（which compsed of poriacocos, alisma rhizoma, atractylodes） throughcomparing DSS with its decompositions, so as to clarify the compatibility of science. Inorder to provide experimental basis for speeding up DSS applied in treatment of livercirrhosis ascites. We study the mechanism of DSS on liver cirrhosis ascites, through theapproach the arginine vasopressin （AVP） and aquaporin.Methods:①T hemethod of phenobarbital combined CCl4is adopted to establish the ratmodel of cirrhotic ascites.②Metabolic cages were used to collect urine in24hours, formeasuring the urine volume. The filter paper was used to absorb ascites.③The levelsof serum ALT, AST, D-BIL, ALB, BUN, Cr were detected by the automaticbiochemistry analyzer. PTwas evaluated by the blood coagulation analyzer.④The indexof liver fibrosis HA, Ln, PCⅢ, CⅣ were determined with the radioimmunoassay（RIA）.⑤The Liver pathological changes were observed by HE staining.⑥The level of K⁺,Na⁺, Cl^-were detected by the automatic biochemistry analyzer.⑦The plasma level ofAVP, AngⅡ, ET-1were detected by enzyme-linked immunosorbent assay （ELISA）.⑧Urine osmotic pressure was detected by cryoscopy; AQP2concentration of urine wasdetected by ELISA.⑨The immunohistochemistry was used to detect the expression ofAQP4on liver and V2R and AQP2on kidney.⑩The expression of V2R-mRNA andAQP2-mRNA were detected by RT-PCR. The protein expression of V2R and AQP2 were detected by Western Blotting.Result：①The reproducible and stable model of the liver cirrhosis ascites rats wereinduced by phenobarbital solution for one week, then intraperitoneal injection of CCl4oil solution for13weeks.②Compared with the model group, urine volume significantlyincreased in24hours and the amount of ascites decreased differently in groups of DSS.③The liver function level of serum ALT, AST, D-BIL, and the kidney function of serumBUN, Cr were significantly decreased; the level of serum ALB increased greatly.Meanwhile, the time of PT was shortened significantly.④The index of liver fibrosisHA, Ln, PCⅢ, CⅣ decreased differently in groups.⑤The groups of DSS recordednotable decreased in fatty degeneration and inflammatory necrosis in liver cells. Thedegree of fibrous septums and pseudo lobules had significantly reduced.⑥The serumlevel of K⁺, Na⁺, Cl^-increased greatly, while the urine level of K⁺, Na⁺, Cl^-decreasedsignificantly.⑦The plasma level of AVP, AngⅡ, ET-1significantly reduced in DSSgroups.⑧Urine osmotic pressure decreased and the level of urine AQP2increasedgreatly.⑨The expression of AQP4increased on bile duct cells of liver, the expressionof V2R and AQP2on kidney decreased significantly.⑩The expression of V2R-mRNAand AQP2-mRNA reduced clearly, meanwhile, the protein expression of V2R andAQP2descend in DSS groups.Conclusion: The stable model of the liver cirrhosis ascites rats could reproduced by themethod of phenobarbital conbined CCl₄oil solution.The prevention effects ofDanggui-Shaoyao-San on cirrhosis rats with ascites include improving hepatic and renalfunction, retarding the liver fibrosis and cirrhosis pathological change and decreasingthe ascites generation. which may be related to its influence on the AVP system andAQPs.