| Objective: To investigate the effect of Yindan Xinnaotong capsule(YDXNT)on carotid intimal hyperplasia caused by balloon injury in rats,and to explore whether the mechanism is related to Wnt/β-catenin and Ca N/NFAT signaling pathway.Methods: SD rats were randomly divided into Sham group(Sham,n = 10)and the carotid artery intimal hyperplasia group,which was established by balloon-injured,all the carotid artery intimal hyperplasia of rats were randomly divided into Model group(Model,n = 10)and YDXNT group(1.0 g/kg,n = 10).YDXNT group was intragastrically administered YDXNT(1.0 g/kg)for 14 days.The Sham and Model groups were intragastrically administered the equal volume of double distilled water.After 14 days of administration,the intimal hyperplasia of the carotid artery was evaluated by small animal ultrasound and the intimal thickness was calculated.The pathological changes of the carotid artery were observed by H&E staining and TUNEL staining.The protein levels of Wnt3 a,β-catenin and PCNA were detected by Western Blot in carotid artery tissues of SD rats.The protein expression of PCNA,α-SMA,OPN,MMP-9,TIMP-1,Bcl-2,Bax,Cyt C,Caspase 9,Caspase 3,Wnt3 a,β-catenin and GSK-3β were detected by immunohistochemical staining in carotid artery neointima of SD rats.The protein expression of IL-1β,TNF-α,MCP-1,Ca N and NFAT4 were detected by immunofluorescence staining in carotid artery neointima of SD rats.Results: Compared with the Sham group,the thickness of the carotid artery in the Model group was significantly increased 68.48%(P<0.05);the neointimal area(NIA),the ratio of neointimal area to media area(NIA/MA),and the ratio of neointimal area to internal elastic lamina area(NIA/IELA)were increased significantly(P<0.05);the cell apoptosis rate of the carotid artery neointimal was increased(P<0.05);Western Blot suggested the protein levels of Wnt3 a,β-catenin and PCNA were significantly up-regulated(P<0.05);immunohistochemical staining showed that the protein expressions of PCNA,OPN,MMP-9,Bcl-2,Wnt3 a and β-catenin were increased(P<0.05),the protein expressions ofα-SMA,TIMP-1,Bax,Caspase 9,Caspase 3,Cyt C and GSK-3β were increased(P<0.05);immunofluorescence staining demonstrated that the protein expressions of IL-1β,TNF-α,MCP-1,Ca N and NFAT4 were increased.Compared with the Model group,the thickness of the carotid artery in the YDXNT group was significantly reduced 18.95%(P<0.05);the neointimal area(NIA)was significantly decreased 77.37%(P < 0.05),the ratio of neointimal area to media area(NIA/MA)was significantly decreased 42.14%(P<0.05),and the ratio of neointimal area to internal elastic lamina area(NIA/IELA)was significantly decreased 36.64%(P < 0.05);the cell apoptosis rate of the carotid artery neointimal was increased significantly(P < 0.05);Western Blot suggested the protein levels of Wnt3 a,β-catenin and PCNA were significantly down-regulated(P < 0.05);immunohistochemical staining showed that the protein expressions of PCNA,OPN,MMP-9,Bcl-2,Wnt3 a and β-catenin were decreased(P<0.05),the protein expressions ofα-SMA,TIMP-1,Bax,Caspase 9,Caspase 3,Cyt C and GSK-3β were increased significantly(P < 0.05);immunofluorescence staining demonstrated that the protein expressions of IL-1β,TNF-α,MCP-1,Ca N and NFAT4 were decreased.Conclusion:(1)YDXNT can improve carotid intimal hyperplasia caused by balloon injury in rats;(2)YDXNT can improve carotid intimal hyperplasia caused by balloon injury in rats by inhibiting the proliferation,phenotypic switching and migration of vascular smooth muscle cell(VSMC),promoting the apoptosis of VSMC,reducing local inflammation of blood vessels;(3)YDXNT may improve carotid intimal hyperplasia caused by balloon injury in rats via inhibiting Wnt3a/β-catenin signaling pathway;(4)YDXNT may improve carotid intimal hyperplasia caused by balloon injury in rats via inhibiting Ca N/NFAT signaling pathway. |
