Study On The Effect Of Fuzi Lizhong Decocion Of AQP4 And ANP In Rats With Spleen Yang Deficiency Syndrome And Its Intervention Mechanism

Objective:To observe the effect of Fuzi Lizhong decoction on content o f aquaporin 4（AQP4）and atrial natriuretic peptide（ANP）in rats with Sp-leen Yang deficiency syndrome,and discuss the mechanism of water m etabolism regulation in rats with Spleen Yang deficiency syndrome.Methods:120 clean grade Wistar rats,both female and male rats,were randomly divided into blank control group,sham operation group,model group,Fuzi Lizhong decoction low-dose group,middle-dose group and high-dose group 6 groups with 20 rats in each group.One hundred of them were administrated by incising brown adipose tissue（BAT）,high fatty diet and cold enviroment stimulating,to establish the animal model of spleen yang deficiency,BAT was found in sham operated group but not incised.The control group was given ordinary diet,and from the first day after incised BAT,the other groups was given high fatty diet for 30 days,and low-dose group,middle-dose group and high-dose group were ig respectively with Fuzi Lizhong decoction(10,20,40 g·kg^-1),the control group and model group were given equal volume of normal salineonce,a day for 30 days.During this period,animal behavior was observed every day,The weight changes of rats were recorded regularly.30 days later,the model group was on the first day after the modeling sucessed,draw all materials that we need.The levels of serum creatinine（SCr）,urea nitrogen（BUN）and urinary creatinine（UCr）were measured by routine biochemical methods;the mRNA expression of AQP4 in rat gastrointestinal tissue were measured by fluorescent quantitative Real Time-Polymerase Chain Reaction（RT-PCR）;detection of AQP4 content in rat gastrointestinal tissue by Western blot;examine content of ANP by ELISA.What’s more,The data were expressed as mean±standard deviation（±S）,and the comparison between two groups were realized by statistical analysis of variance,the difference has statistical significance（P<0.05）.Results:General observation:The rats of model group were depressed,cold chills,curled,extrados,lesslively,haggardfur,and anorexia and loss of weight,ilassitude,loos stools and perianal contamination etc.2 weeks after the treatment,all the above phenomenons and symptoms were improved in different degree.Under the light microscope observation showed that the pathological changes of HE staining of rat gastrointestinal tissue were gastrointestinal tissue in rats alleviated in varying degrees,and the high dose group had the best effect.Comparing with the model group,The results of RT-PCR showed that the expression of AQP4 mRNA in middle and high dose group was relatively high,and the difference was statistically significant（P<0.05,P<0.01）;The results of Western blot showed that the content of AQP4in middle and high dose group was relatively high,the difference was statistically significant（P<0.05,P<0.01）.The results of ELISA showed that the content of ANP in middle and high dose group was relatively high,the difference was statistically significant（P<0.05,P<0.01）.Conclusions:Fuzilizhong decoction can Improve the content of AQP4,ANP and AQP4 mRNA and relieve diarrhea symptoms of Spleen Yang Deficiency Rats.The degree of improvement was positively correlated with the dosage.AQP4 and ANP are both likely to be the molecular biological basis in regulating body fluid metabolism in rats with Spleen Yang deficiency syndrome with Fuzilizhong decoction.