Relative Studies On The Modulation Of NO And ET-1 In Cirrhosis With Ascites Treated With Formula Of Invigorating Spleen And Tonifying Liver

Objectives:"Five viscera correlation" theory is a clinical theory of Traditional Chinese medicine(TCM)that has been advocated by famous veteran doctor of TCM-Prof.Deng Tietao for years.As one of Pro.Deng' s students, my advisor Prof.Liu Youzhang inherited systematically his academic ideology, and expanded the "liver-spleen correlation" theory which is an important branch of "five viscera correlation" theory."Liver-spleen correlation" theory has profuse substantial connotations,which central contents expound the physiological functions,pathological changes,syndromes,treatments and their corresponding formulas.According to this theory,liver-correlative diseases which locate in the liver,have intimate correlations with the spleen. Cirrhosis with ascites is one of the common and refractory diseases,which belongs to "flank pain","accumulation","drum belly" in TCM and jeopardizes the patients actuely.Clinically support treatments are mainly used,but the curative effect is common.My advisor reckons that hepatic cirrhosis results from the liver failing to maintain the normal flow of qi due to various reasons,liver and spleen comorbidity,deficiency and blood stasis due to invalidism.So the basic pathogenesis of hepatic cirrhosis is spleen deficiency and blood stasis.Ultimately it appears as body fluid and pathogenic damp detention.My advisor has been treated cirrhosis with ascites with formula of invigorating spleen and tonifying liver(FISATL)which is composed of characteristic Lingnan Chinese herbal medicine for years,and has achieved significant curative effect.This project is to further verify the curative effect of FISATL through clinical observation and animal experiment, and approach the mechanism of action of FISATL through contrast observation of relative indexes,and also provide relative experimental data about the effectiveness and scientificalness in treatment of cirrhosis with ascites. Methods:This study includes literature research,experimental research.In literature research,the apprehension of TCM for liver and cirrhosis with ascites(drum belly)in the ancient times,and the studies of modern medicine of the relations between cirrhosis with ascites,portal hypertension and vasoactive substances,and the mechanisms how cirrhosis with ascites occurs and animal modeling were all arranged and summarized in detail.Experimental research is composed of two parts.In the first part,according to visit order, 40 cases of cirrhosis with ascites according with spleen deficiency-damp abundance- blood stasis syndrome,were divided randomly into control group and thera- peutic group。Each group had 20 cases.Control group received elementary treatment of western medicine.Therapeutic group added decoction of FISATL,one dose per day,in addition to the treatment of control group. Those treat- ments went on for 3 months.Abdominal girth,clinical symptoms, hepatic function,ultrasonograph of liver and spleen,NO and ET-1 from peripheral blood were all observed before and after the treatment.In the second part,90 Wistar rats which were specific pathogen free(SPF)were selected. 10 rats selected randomly became blank group(group A).The rest were injected subcutaneous with 40%carbon tetrachloride(CCL₄)dissolved in olive oil. These rats were fed on liquor as their only hydroposia.After 12 weeks of molding for cirrhosis with ascites,54 rats were left.Conformed by diagnostic peritoneocentesises,44 rats with ascites were divided into 5 groups which included model group(group B),FISATL large dose,medium dose,small dose group(group C,D,E)and began a 4-week intragastric administration.Each group had 9 rats,and the colchicine group(group F)had 8 rats.Group C,D, E were given FISATL respectively in the dose of 52g/kg body weight(BW), 26g/kg BW,16.25g/kg BW.group F was given colchicine 100μg/kg BW.The intragastricalamount was 10ml/kg BW.Administration was given once per day, the concentration and equivalent dose were converted according to BW.At the end of the experiment,general state of health,body weight and the amount of ascites of those rats were observed.Pathology changes of the hepatic tissues after HE and VG stain were observed.Portal venous pressure was measured.The indexes of hepatic function,NO,and ET-1 were detected from portal vein blood of those rats.Result:1.The total effective rate in therapeutic group was 85%,50%in control group. Therapeutic group outpassed control group.After statistical treatment,the differences was significant(p＜0.05).1.1 Abdominal girth comparison before and after the treatment:the differences were significant(p＜0.01).The diminution degree of the therapeutic group was greater than the control group,and the differences were significant(p＜0.05).1.2 Symptoms comparison before and after the treatment:There was obviously improvement in lack of appetite,fatigue,abdominal distension pain in flank in therapeutic group(p＜0.01),also improvement in the texture of tongue(p＜0.05).The improvement in control group was slower(p＞0.05).1.3 Comparison of indexes of liver function before and after treatment: the contents of ALT,AST of both groups and the contents of GLB of therapeutic group all decreased significantly after the treatment(p＜0.01).The contents of ALB of both groups heightened after the treatment,and the differences were significant(p＜0.01).The contents of GLB of control group decreased slightly after the treatment,but there was no significant difference before and after the treatment(P＞0.05).The difference of the contents of GLB between the two group was significant(p＜0.01).1.4 Ultrasonograph comparison before and after the treatment:There was improvement in oblique diameter of right hepatic lobe,thickness of spleen, inside diameter of portal vein and splenic vein after the treatment.But slightly diminished oblique diameter of right hepatic lobe,increment of spleen thickness,widened inside diameter of portal vein and splenic vein were observed in control group after the treatment.The differences between those indexes of the two group were not significant(p＞0.05).1.5 Integral calculus of ascitic capacity in both groups after the treatment all decreased.The differences before and after the treatment were significant in therapeutic group(p＜0.05),but not in control group(p＞0.05). And there were significant differences between the two groups after the treatment,indicating the regression of ascites in therapeutic group was greater than that in control group.1.6 Comparison of the contents of NO and ET-1 in peripheral blood before and after the treatment:The contents of NO and ET-1 both decreased significantly in therapeutic group(p＜0.01),but heightened slightly in control group(p＞0.05).The contents of NO and ET-1 in therapeutic group was much lower than those in control group,and the difference was significant (p＜0.01).1.7 The incidence of complications in therapeutic group was 20%,35%in control group,and the difference was not significant(p＞0.05).There were 4 cases in therapeutic group,and 7 cases in control group.2.General state of health of the rats:the mortality of modeling rats was 32.5%.Ascites formation rate was 55%.It is observed that tangled and bloom fur,descent of activity amount,decrease of drink and food,loss of weight, loose feces occurred in the model group rats with ascites in cirrhosis.After the treatment,all of the changes mentioned above were improved.2.1 After the experiment,there was no ascites in group Aand much ascites in groups B and F,which was canary.The amount of ascites decreased significantly in each FISATL therapeutic group.Compared to group B,the differences were very significant(p＜0.01).2.2 HE and VG stain of liver tissues:FISATL could obviously relieve the inflammation of the liver tissues and the hyperplasia of collagen fibers. The improvement in liver structure of Group C,D were more obvious,which was correspondent to group F(p＞0.05).2.3 The contents of indexes of hepatic function including ALT,AST,GLB heightened obviously in model group rats(compared to groupA,p＜0.01). After the treatment in Group C,D,E,F the contents of ALT,AST,GLB decreased unequally,and the contents of ALB heightened.The effectiveness in group C was the best.Compared with group A,there was no significant differences after the treatment in those indexes(p＞0.05).But there was significant differences between each group(D,E,F)and group A after the treatment(p＜0.01).2.4 Portal venous pressure of the rats of group C(9.42±0.96mmHg) obviously lower portal venous pressure of group B(14.49±1.28mmHg),which was near the pressure of group A(8.22±1.07mmHg)(p＞0.05).Compared to model group,the difference was significant(p＜0.01).Pressure from other therapeutic groups(group D,E,F)dropped too,but the differences compared to group A were significant(p＜0.01).2.5 The contents of NO and ET-1 in portal vein blood of rats with ascites in cirrhosis were much higher than those of group A.After the treatment,the contents of NO and ET-1 in group C,D,E all decreased,and group C had the greatest descent.The contents of NO and ET-1 in group F didn't change obviously(compared to model group,p＞0.05).Conclusions:1.Improved molding method of composite factor is one of the ideal methods to create models with ascites in cirrhosis.2.FISATL could obviously improved the mental state,color pattern,appetite, and body weight of the rats,and diminish the amount of ascites of the rats, indicating good effect on ascites regression.3.FISATL could obviously relieve the inflammation of the liver tissues and the hyperplasia of collagen fibers,which showed better effectiveness on liver protection and anti-hepatic fibrosis.4.FISATL could lower portal venous pressure of the model rats.5.FISATL could lower the contents of NO and ET-1 in portal vein blood, improved portal venous flow and its resistance in the rats with ascites in cirrhosis.This could be the main mechanism which FISATL resists portal hypertension.6.The effectiveness of FISATL contains invigorating spleen and benefiting vital qi,activating blood circulation to dissipate blood stasis,clearing heat and dissipating dampness,and regulating vital qi and stomach.With the appliance of FISATL,clinical symptoms and liver function of patients with ascites in cirrhosis corresponding to spleen deficiency-damp abundance-blood stasis syndrome could be improved significantly,and the contents of NO and ET-1 in peripheral blood could be down regulated.And the appliance of FISATL could also improve the sizes of liver and spleen,inside diameter of portal vein and splenic vein.So the appliance of FISATL is worthwhile to be approached and studied furtherly.