The Effect Of FGC-3 To AQP8?ET-1?NO Of Cirrhotic Ascites Rats

Objective: This experiment adopted an improved CCl4–alcohol method to prepare the rat model with cirrhotic ascites,and then used Fuganchun 3(FGC-3)and Compound Biejiarangan Troche to intervene the model.By observing the rats' general situation,amount of ascites,liver function,liver fibrosis indexes,nitric oxide(NO),endothelin-1(ET-1),aquaporin 8(AQP8),and the pathological changes of liver,to study the effect of FGC-3 on ascites and liver,explore the possible mechanism of FGC-3 on cirrhotic ascites,and wish to provide experimental basis for the clinical treatment of cirrhotic ascites by traditional Chinese medicine.Method: Seventy-five clean male wistar rats,weight 200-220 g,fedadaptability for one week.8 rats was selected randomly as normal control group(N group),and the other rats were made model by subcutaneous injection of 40% CCl4-olive oil,every 4 times,the first dose of 0.5ml/100 g,starting from the second dose to 0.3 ml/100 g,and adjust the dosage according to the weight,combined with 15% ethanol as a free drink and free ordinary feed.After 10 weeks,41 rats had ascites by checking sign of ascites and abdominal diagnostic aspiration,then divided these 41 rats into 5 groups randomly: Model Group(M Group)nine,High dose Group of FGC-3(G Group)eight,Middle dose Group of FGC-3(Z Group)eight,Low dose Group of FGC-3(D Group)eight,Compound Biejiarangan Troche Group(F Group)eight.Each group gave corresponding drug intragastrically everyday for one month.All rats were fasted above 12 hours,then we drew the blood from the rats' eyeball for measuring the blood indicators.After killing the rats by cervical dislocation method,to open its abdominal cavity,and extracted abdominal cavity effusion;To observe the color and texture of rats' liver,then put the rat liver tissue into cryopreserved tubes todetect AQP8,which were preserved in liquid nitrogen at low temperature;The same site liver Tissue was soaked in 4% neutral formaldehyde for HE staining slice.Results:1 Comparison of general condition of the ratsIn the experiment the mortality of the rats who were made model was 20.9%,and the ascites formation rate was 61.19%.The rats of N Group was in good spirits,acted normally;their fur and feather,bite and sup and urine and stool had no noticeable change,and their weight gradually increased.Rats of M Group were depressed obviously,easy to be scared.They had less activity and bite and sup.Their fur and feather were yellow,dried-up and easy to fall off.Their stool were ashen,and their weight first reduced and then increased.After treatment,the above situation of treatment groups had inordinately improvement.2 Comparison of the rats' liver appearanceWith the naked eye observation of rat liver tissue,the liver of N group was of good texture,bright red,smooth surface,sharp edges and moderate size;M group was crisp,color dark red or brown,uneven surface,visible nodules and blunt edge;D group was swelling,crisp,color brown,and its surface was coarse granular,and its edge wss more blunt;And G,Z and F group,hepatomegaly,dark red,sharp edge and their surface were still smooth,but showed different degrees of fine granular area.3 Comparison of the amount of ascitesThe ascites volume of M group increased significantly,compared with the normal group,the difference was statistically significant,P<0.05;after treatment,the ascites of G,Z,D and F group had different degrees of reduction,in which G,Z,D group decreased significantly,the difference is statistically significant,P<0.05;The difference between M group and F Group was not statistically significant,P>0.05.4 Comparison of liver function in different groupsCompared with N group,the other groups' AST and ALT were increased to varying degrees,in which M,F,Z and D group had a significant difference,P<0.05;G group had no significant difference,P>0.05.Compared with M group,each treatment group's AST and ALT were reduced to varying degrees,the difference had statistically significant,P<0.05.Compared with N group,the other groups' ALB were decreased to varying degrees,in which M,F,Z and D group had a significant difference,P<0.05;G group had no significant difference,P>0.05.Compared with M group,each treatment group's ALB were increased to varying degrees,in which G and Z group had statistically significant,P<0.05;F and D group had no statistically significant,P>0.05.5 Comparison of liver fibrosis indexes in different groupsCompared with N group,the liver fibrosis index of other groups were increased to varying degrees,and the difference had statistically significant,P<0.05;Compared with M group,each treatment group was reduced to varying degrees,the difference had statistically significant,P<0.05,and in which G group improved significantly.6 Comparison of NO in different groupsCompared with N group,the NO 0f M group was significantly higher,and the difference was statistically significant,P<0.05.Compared with M group each treatment group was reduced to varying degrees,in which G,Z and D group had statistically difference,P<0.05;F group had no statistically difference,P>0.05.7 Comparison of ET-1 in different groupsCompared with N group,the ET-1 0f M group was significantly higher,and the difference was statistically significant,P<0.05.Compared with M group each treatment group was reduced to varying degrees,in which G,Z and D group had statistically difference,P<0.05;F group had no statistically difference,P>0.05.8 Comparison of AQP8 in different groupsThe results of immunohistochemistry showed that brown granules could be seen around the liver cells of rats N,which suggested the expression of AQP8 in the region.In comparation,the expression of AQP8 in M group was decreased significantly,and the difference was statistically significant,P<0.05.Compared with M group,G and Z group had significant difference,P<0.05;F and D group had no statistically difference,P>0.05.Image analysis of the data showed that the liver AQP8 integral optical density(IOD)of M group was lower than that in N group,and the difference was statistically significant,P<0.05.Compared with M group,the IOD of G and Z group significantly increased,the difference was statistically significant,P<0.05;F and D group increased slightly,but the difference was not statistically significant,P>0.05.Conclusions:1 The improved CCL4-alcohol comprehensive method can make a stable liver cirrhosis ascites model in rats.2 FGC-3 can obviously improve the general situation of liver cirrhosis ascites rats,significantly decrease the amount of ascites,and promote the ascites subsided.3 FGC-3 can significantly reduce the AST,ALT and liver fibrosis index level,elevate the level of ALB,reduce the degree of fibrosis and inflammation of the liver,and has good effect of liver protection and anti fibrosis.4 FGC-3 can decrease the levels of NO and ET-1,thereby improves the hyperdynamic circulatory state,and then reduces portal pressure,which may be one of the mechanisms of cirrhosis ascites.5 FGC-3 can enhance the expression of AQP8 in cirrhotic rats with ascites,which may be one of the mechanisms of cirrhosis ascites.