| Portal hypertension(PH)is a common and refractory complication of chronic liver disease.It is characterized by increased blood flow and increased resistance of portal vein,lacking of specific treatment drugs.At the same time,the activity of superoxide dismutase 3(SOD3)and the bioavailability of nitric oxide(NO)and prostacyclin(PGI2)decreased,which were the main factors of portal hypertension.In the early stage of this project,we established a constant flow pressure rat portal vein perfusion system in vitro,and found that Chinese medicine monomer diammonium glycyrrhizinate and salvianolic acid B could effectively reduce portal vein pressure and relieve PH.Therefore,on the basis of previous studies,the PH model of rats was induced by carbon tetrachloride(CCl4).The macrophages in the liver were removed by liposomes of disodium chlorophosphonate,and the activation of macrophages was determined to participate in PH.In vitro perfusion portal system,aminoguanidine hydrochloride inhibited iNOS,celecoxib inhibited COX-2,observed the bioavailability of diastolic substance,and explored the pharmacological mechanism of diammonium glycyrrhizinate and salvianolic acid B in the prevention and treatment of pH.1.Macrophages participate in portal hypertension in rats[Objective]To observe the effect of macrophage activation on portal hypertension in rats.[Methods]Forty male Wistar rats were randomly divided into two groups:the control group received subcutaneous injection of olive oil twice a week(3mL/kg),the model group received subcutaneous injection of 40%CC14 twice a week(3mL/kg);d70-105,the model rats were divided into two subgroups,the macrophages were not removed and cleared group,half of the animals received caudal injection of PBS liposome once a week(0.3mL/kg),the other half of the animals were injected with liposome of disodium chlorophosphonate(15mg/kg).d105.Blood was collected from abdominal aorta of rats in each group.The number of monocytes in plasma,the content of CD 163 and CCL2 in serum were measured.The left lobe liver was taken and the expression of CD 163 in liver was detected by immunohistochemistry.[Results]In the PBS liposome group,compared with the control group,the number and percentage of monocytes in the model group increased,the pressure of portal vein increased,the content of serum CD 163 and CCL2 increased,and the expression of CD 163 in the portal area increased;in the macrophage dealt groups,compared with the control group,the number and percentage of monocytes in the model group increased,Compared with the control group,the number and percentage of monocytes in the macrophage clearance group decreased,the content of CD 163 and CCL2 in the serum decreased,the content of CD 163 and CCL2 in the serum decreased,In the model rats,the number and percentage of monocytes,portal pressure,sCD163,CCL2 and CD 163 expression in the macrophage clearance group were lower than those in the non clearance group.[Conclusion]Macrophages participate in portal hypertension in rats.2.Effect of Yiqi-huoxue prescription active ingredients on prevention and treatment of portal hypertension in rats[Objective]To observe the effect of diammonium glycyrrhizinate and salvianolic acid B on portal hypertension in rats.[Methods]132 male Wistar rats were divided into control group,model group,positive group,salvianolic acid B group,diammonium glycyrrhizinate group and combined group.Olive oil(3mL/kg)was injected subcutaneously twice a week in the control group,40%CCl4(3mL/kg)was injected subcutaneously twice a week in the model group and the treatment groups.On d56,the control group and the model group were given intragastric gavage every day 0.01%CMC(5mL/kg),Each treatment group was given captopril,salvianolic acid B,diammonium glycyrrhizinate,salvianolic acid B and diammonium glycyrrhizinate(5mL/kg)by gavage on d70.The rats in each group were divided into two subgroups.Liposomes were injected into tail vein once a week(0.3mL/kg).After anesthesia,ascites was collected and the pressure of portal vein was measured,the weight of liver and spleen were measured.Hematoxylin eosin(HE)and Masson staining were used to observe the pathological changes of liver and collagen deposition.The number and volume ratio of hepatic lobules(pseudolobules)and the ratio of collagen fiber were measured.[Results]Compared with the model group,the positive drug captopril and the single Chinese medicine group could significantly reduce the amount of ascites,alleviate the pathological changes of the liver,reduce the formation of pseudolobules and collagen deposition,reduce the pressure of portal vein in vivo,and improve the liver function;after macrophage removal,it was compared with the control group,The number of pseudolobules decreased,the average volume increased and the deposition ratio of collagen fibers decreased in the macrophage removal group.[Conclusion]Salvianolic acid B and diammonium glycyrrhizinate can effectively treat portal hypertension in rats,and the therapeutic effect is macrophage dependent.3.Pharmacology of Yiqi-huoxue prescription active ingredients inhibiting macrophage hyperoxia and preventing portal hypertension[Objective]To explore the pharmacological mechanism of salvianolic acid B and diammonium glycyrrhizinate in inhibiting macrophage hyperoxia in portal area and preventing portal hypertension in rats.[Methods]144 male Wistar rats were divided into control group,model group,positive group,salvianolic acid B group,diammonium glycyrrhizinate group and combined group.Olive oil(3mL/kg)was injected subcutaneously twice a week in the control group,40%CCl4(3mL/kg)was injected subcutaneously twice a week in the model group and the treatment groups.On d56,the control group and the model group were given intragastric gavage every day 0.01%CMC(5mL/kg),Each treatment group was given captopril,salvianolic acid B,diammonium glycyrrhizinate,two monomers of the same amount of mixture(5mL/kg)gavage treatment,d70,the rats in each group were divided into two subgroups,macrophages were not cleared and cleared group,PBS liposomes were injected into tail vein once a week(0.3mL/kg),the other half of the animals were injected with chlorophosphonate liposome(15mg/kg)intravenously.We measured the portal pressure in vivo,perfused the portal vein in vitro,measured the utilization of NO and PGI2,took the left lobe liver,measured the positive expression of NADPH oxidase 2(NOX2),SOD3,iNOS,inducible nitric oxide synthetase(eNOS),liver homogenate,detected the activity of SOD3,the expression of NOX2 and SOD3.[Results]Compared with the model group,the pressure of portal vein decreased,the expression of NOX2 and iNOS decreased,the expression of eNOS and SOD3 increased,the activity of SOD3 increased,the utilization of diastolic NO and PGI2 increased;after macrophage clearance,compared with the control group,the pressure of portal vein decreased,the expression of NOX2 and iNOS decreased,the expression of eNOS and SOD3 increased,the activity of SOD3 increased,the utilization of diastolic NO and PGI2 increased.The portal vein pressure decreased,the expression of NOX2,iNOS and eNOS decreased,the utilization of NO and PGI2 increased in salvianolic acid B and combination group,and the utilization of NO and PGI2 decreased in diammonium glycyrrhizinate group[Conclusion]Macrophage is one of the targets of salvianolic acid B in the treatment of portal hypertension.Salvianolic acid B and diammonium glycyrrhizinater can improve the utilization of diastolic substance,protect the activity of antioxidant enzymes,reduce portal hypertension and prevent portal hypertension. |
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