| Objective:1.To explore the risk factors and TCM syndrome characteristics of cirrhosiscomplicated with portal vein thrombosis.2.The model of rat model of cirrhotic portal vein thrombosis was established by usingthe main part of the portal vein to ligature and compound CCl4 intraperitoneal injection.3.To observe the therapeutic effect and dose-effect relationship of Danggui Buxue Decoction on animal models of cirrhotic portal vein rats.Method:1.To collect patients with liver cirrhosis in the second ward of the Department of Hepatology,Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,according to the corresponding standard,collect relevant medical history data,laboratory and auxiliary examination reports,and the control group was included in the same level of cirrhosis patients with Child score.Establish a clinical information database.The clinical data of the two groups were compared to analyze the risk factors and independent risk factors of cirrhosis complicated with portal vein thrombosis.The clinical characteristics of patients with cirrhosis complicated with portal vein thrombosis were analyzed.2.A rat model of portal cirrhosis with portal vein thrombosis was induced by ligation of the main part of the portal vein combined with CC14 intraperitoneal injection.Observe and compare the general conditions of 4,6,8 and 10 weeks of dynamic time in rat animal models,including changes in liver/body ratio,spleen/body ratio;HE staining of liver tissue and detection of serum ALT and AST To observe the level of liver tissue inflammation;Sirius red staining and hydroxyproline content determination,observation of liver tissue collagen deposition and liver fibrosis,cirrhosis;serum prothrombin time,INR,prothrombin activity,activated partial clotting activity Enzyme time,fibrinogen,D-dimer,antithrombin ? and platelet were detected to evaluate the changes of coagulation function in rat animal model;ultrasonic method was used to detect the diameter and blood flow velocity of portal vein in rat model;After anatomy,rat portal vein,HE staining of portal vein and Fibrinogen immunohistochemical staining of liver tissue were used to evaluate portal vein thrombosis in rat model.3.The therapeutic effect,dose-effect relationship and mechanism of Danggui Buxue Decoction on the animal model of cirrhotic portal vein thrombosis induced by ligation of the main part of the portal vein combined with CCl4 intraperitoneal injection.Compare normal group,model group,Danggui Buxue Decoction low,high dose group and positive drug control group animal model,including body weight,liver/body ratio,spleen/body ratio changes;liver tissue HE staining and serum ALT,AST detection,observation of changes in liver tissue inflammation;Sirius red staining,observation of liver tissue collagen deposition and liver fibrosis,liver cirrhosis improvement;serum prothrombin time,INR,prothrombin activity,activation Detection of thromboplastin time,fibrinogen,D-dimer,antithrombin III and platelets,assessment of changes in coagulation function in rat animal models;ultrasound method for detection of portal vein diameter and blood flow velocity in rat animal models Changes;immunofluorescence and Western blotting were used to detect the expression of vWF,eNOS,Rab3B and Rab27A in liver tissue,and to evaluate endothelial cell function.Result:1.126 patients with cirrhosis were collected,including 62 patients with portal vein thrombosis.Univariate analysis showed that patients with concurrent portal vein thrombosis and no portal vein thrombosis had cholinesterase,fibrin degradation products,D-dimer,neutrophil percentage,portal vein diameter,portal vein flow velocity,spleen vein diameter,The differences in spleen thickness,spleen long diameter,and peritoneal effusion were statistically significant(P<0.05).Logistic regression analysis showed that D-dimer(OR=2.078,95%CI 1.293-3.339,P=0.003),spleen long diameter(OR=1.964,95%CI 1.228-3.142,P=0.005)is an independent risk factor for cirrhosis with portal vein thrombosis;fatigue,splenomegaly,bloating,severe esophageal varices,variceal hemorrhage History,pale tongue,pulse string/fine is the main feature of patients with cirrhosis complicated with PVT,consistent with the performance of qi deficiency and blood stasis syndrome2.Dynamic time point observation of rat model of cirrhotic portal vein thrombosis induced by ligation of the main part of the portal vein combined with CC14 intraperitoneal injection.Biochemical results:serum ALT and AST were significantly increased in each composite model;the 10-week model group was increased in different degrees compared with each model group.The hydroxyproline content in the liver tissue of each composite model gradually increased,and the severity of liver fibrosis gradually increased.The 10-week model group was the most significant.HE staining of liver tissue:bile duct hyperplasia in the portal area of the simple ligation group.In the composite model,the liver tissue was formed with the prolongation of the modeling time.At the 10th week,the fiber spacing increased and thickened.A large number of bile duct epithelial cells and inflammatory cells infiltrated,and granulomas were formed.Sirius red staining:thickening of the central venous wall of the simple ligation group.In the composite model group,with the prolongation of modeling time,the collagen in the central venous wall gradually thickened,and the deposition of collagen fibers in the portal area gradually increased.The combination of the portal area and the portal area appeared,and the 10-week model showed collagen fiber wrapping.The agglomerates form a plurality of fiber spaces.Prothrombin time:6 weeks and 10 weeks model significantly increased;INR:6 weeks and 10 weeks model decreased significantly;prothrombin activity:significantly decreased in each model group.Activated partial thromboplastin time:The 10-week model was significantly elevated.Fibrinogen:A significant decrease in the 10-week model group.D-dimer:A significant decrease in the 10-week model group.Antithrombin III:Significant decline in the model group at 4 weeks,6 weeks,and 10 weeks.Platelets:The 6-week model group decreased,and the remaining model groups all showed a downward trend.Inner diameter and flow rate of portal vein:The inner diameter of the main group of the model group was significantly narrowed at 10 weeks;the flow rate of each composite model group was significantly slowed down.3.The efficacy and dose-effect relationship of Danggui Buxue Decoction in the rat model of cirrhotic portal vein thrombosis induced by ligation of the main part of the portal vein combined with CC14 intraperitoneal injection.Biochemical results:ALT decreased in the high-dose group of Danggui Buxue Decoction group and rivaroxaban group.The hydroxyproline content of Danggui Buxue Decoction high dose group and positive drug control group decreased,and Danggui Buxue Decoction high dose group had the largest decrease.HE staining of liver tissue:The inflammatory reaction of Danggui Buxue Decoction group was significantly relieved,no obvious bile duct hyperplasia reaction,central venous lumen was significantly reduced,tube wall thickening was significantly improved;rivarshaban group inflammation and bile duct hyperplasia were alleviated.Sirius red staining:There was no significant reduction in fibrotic spacer collagen deposition in the low-dose group of Danggui Buxue Decoction.The fibrosis was significantly alleviated in the high-dose group,the fiber spacing was incomplete,and the fibrosis collagen deposition in the rivaroxaban group was alleviated.Prothrombin time:Significantly shortened in each treatment group.INR:Significantly shortened in each treatment group.Prothrombin activity:Significantly elevated in each treatment group.The time of activation of partial thromboplastin was lower in the rivaroxaban group than in the low-dose group of Danggui Buxue Decoction and the high-dose group of Danggui Buxue Decoction.Fibrinogen:There is a decreasing trend in each treatment group.D-dimer:There is a decreasing trend in each treatment group.Antithrombin III:Danggui Buxue Decoction high dose group,rivaroxaban group significantly increased.Platelets:There is an increasing trend in each treatment group.Inner diameter and flow rate of portal vein:The inner diameter of each treatment group was significantly widened,and the flow rate of Danggui Buxue Tang low dose group was increased.The results of immunofluorescence showed that the expression of vWF in liver tissue of each treatment group decreased significantly,and the lower dose group of Danggui Buxue Decoction group was lower.The results of Western blot showed that the expression of eNOS in liver tissue of Danggui Buxue Decoction high-dose group and rivaroxaban group increased significantly.The expression of Rab3B in liver tissue of each treatment group was significantly decreased.The lower dose group of Danggui Buxue Decoction group was significantly lower.Compared with the low-dose group of Danggui Buxue Decoction,the expression of Rab27A in the liver tissue of the high-dose group was decreased.onclusion:1.D-dimer,spleen long diameter is an independent risk factor for cirrhotic portal vein thrombosis;patients with cirrhotic portal vein thrombosis meet the characteristics of"qi deficiency and blood stasis" syndrome.2.Rat animal model induced by ligation of the main part of the portal vein combined with CCl4 intraperitoneal injection..Cirrhosis and portal vein thrombosis occurred in CCl4 for 10 weeks.3.Danggui Buxue Decoction has the effects of reducing intrahepatic inflammatory response,anti-fibrosis,inhibition of coagulation function and thrombus dissolution,and the high dose group is superior to the low dose group,which may be related to the protection of endothelial cell injury. |
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