The Role Of Arterial Remodeling In Portal Hypertension

Part?.SMA remodeling in PHTAim:Explore whether SMA remodeling exist in cirrhotic and non-cirrhotic PHT.Methods:Establish CCl₄ inhalation induced cirrhotic and PPVL induced non-cirrhotic PHT rat model,and check condition of model then check SMA remodeling phenotype.Results:MAP were decreased while PP were elevated in two kinds of PHT modeling.Liver gross observation,HE,Masson and Sirius Red staining showed liver cirrhosis changes in cirrhotic PHT,but there were no difference between non-cirrhotic PHT and control rats.SMA thinning occurred in cirrhotic PHT rats but not in non-cirrhotic PHT rats.Conclusion:Both models were successful.A significant arterial remodeling phenotype existed in cirrhotic PHT model,which did not exist in the non-cirrhotic PHT model.Part?.Vascular structural changes in PHT arterial remodelingAim:To elucidate the changes of main components in cirrhotic PHT arterial remodeling.Methods:The expression levels of main structural-related proteins in SMA were detected by electron microscopy,WB,immunohistochemistry and fluorescence.Results:The inner surface of SMA in cirrhotic PHT rats was uneven and EC was shed,while p-e NOS expression increased,and calmodulin,elastin and?-SMA expression decreased.Conclusion:The main changes in SMA remodeling were the endothelial layer shedding,increased basement membrane space,and decreased contractile proteins such as calmodulin,elastin,and?-SMA,while the expression of p-e NOS associated with vasodilation increased.Part ?.VEGFR-3 expression and PHT arterial remodelingAim:To study the role of VEGFR-3 in vascular remodeling of SMA.Methods:The expression of SMA-related genes and proteins in cirrhotic PHT model was detected by PCR and immunofluorescence.Results:The expression of VEGFR-3 on SMA increased in rats with cirrhotic PHT,while the proliferation signaling represented by Ki-67 and the apoptosis signal represented by cleaved caspase-3 increased.Conclusion:In the SMA arterial remodeling of PHT in liver cirrhosis,the expression of VEGFR-3 signal increased,and the expression of its ligand VEGF-D also increased.Vascular proliferation and apoptotic signaling were increased,and the smooth muscle cell layer is dominated by apoptosis.Part ?.Effect of VEGFR-3 inhibition in PHT arterial remodeling Aim:To prove whether inhibition of VEGFR-3 signaling pathway can improve SMA thinning in PHT.Methods:The SMA phenotype was measured after treatment with the VEGFR-3 inhibitor MAZ-51 in cirrhotic PHT rats.Results:There was no significant difference in PP,MAP and VEGFR-3 expression in two groups.MAZ-51 improved SMA remodeling,increased wall thickness and wall area,and reduced SMA smooth muscle layer apoptosis in cirrhotic PHT rats.Conclusion:The treatment of the VEGFR-3 inhibitor MAZ-51 could alleviate SMA arterial remodeling and reduce apoptosis of SMA smooth muscle layer in cirrhotic PHT.Part ?.VEGFR-3 signaling pathway in PHT arterial remodeling Aim:To explore the VEGFR-3 signaling pathway in the SMA EC layer.Methods:The primary isolation of mesenteric arterial EC was used,to stimulate EC with different time and concentration of VEGF-D,and MAZ-51 combined with VEGF-D stimulated EC.Results:With the increase of VEGF-D stimulation time and concentration,the expression of VEGFR-3downstream such as phosphorylated e NOS,AKT and ERK increased gradually,reaching the highest peak at 10 min and 50 ng/ml.Compared with the VEGF-D group,the MAZ-51 group significantly reduced the phosphorylation of downstream proteins AKT,ERK and e NOS.Conclusion:The treatment of the VEGFR-3 inhibitor MAZ-51 can reduce the expression of phosphorylated AKT,ERK,and e NOS,which were downstream of VEGFR-3 signaling in the mesenteric arterial EC.