| Objective Atherosclerosis(AS)is the pathological basis of cardiovascular diseases.Panax notoginseng saponins(PNS),the main component of traditional Chinese medicine Panax notoginseng,can regulate blood lipids,inhibit inflammation,and interfere with the development of AS.20(S)-protopanaxadiol(PPD)is a metabolite of PNS.Caveolae/Cav1 plays an important role in maintaining cholesterol homeostasis,cell signaling,cell proliferation and migration.This study explored the anti-AS mechanisms of PNS and PPD on apolipoprotein E knockout(Apo E-/-)mice and human umbilical vein endothelial cells(HUVECs)through studying the regulation of caveolae/Cav1 in the development of AS.Part ? Effects of PNS and PPD on AS-related lesions in Apo E-/-miceMethod 1.Establishment of animal model:The Apo E-/-mice were fed a normal diet for 6 months and 9 months to establish a AS mouse model.2.PNS and PPD interference:Animals were randomly divided into 6-month-old control group,PNS group(100mg/Kg)and PPD group(20mg/Kg),9-month-old control group,PNS group(100mg/Kg)and PPD group(20mg/Kg).The PNS interference group and the PPD interference group were intraperitoneally injected according to the dose,and the control group was injected with an equal volume of saline once every other day for 3 months.3.Specimen collection and detection:The deposition of vascular lipid plaque was observed.Serum lipid levels were detected.Aortic sinus stenosis and plaque were observed.Peripheral and cardiac microcirculation perfusions were detected.Serum VEGF levels were detected.Results Lipid plaque deposition could be reduced in the 6-month-old and 9-month-old PPD group.Blood lipids could be improved in different degrees in each drug group.Plaque progression could be delayed in each drug group,and stenosis could be improved in the 6-month-old PNS group.The microcirculation of the outer auricle and heart was improved in the 6-month-old and 9-month-old PNS groups,respectively.Serum VEGF was reduced in the 6-month-old PPD group.Conclusion PNS and PPD can resist AS in varying degrees by improving blood lipids,reducing lipid deposition,delaying plaque progression,and improving microcirculation blood flow,and the effects of PPD are better than PNS.Part ? Mechanism of PNS and PPD on HUVECsMethod 1.Cell culture:HUVECs were used as the research objects,cells were cultured with 10%fetal bovine serum.2.Sample collection and testing:Cells were treated according to grouping and medication time and samples were collected.The viability of each group was detected.The structure of caveolae was observed by transmission electron microscopy.The localization and expression of GM1,Cavl and pCav1 proteins were observed.The expression of total protein and plasma protein was detected.The intracellular calcium concentration was detected.The mRNA levels of Cav1,VEGF,ICAM-1 and oxLDLR were detected.Results The best concentrations of PPD,PNS,M?CD were determined.Cell viability was decreased after M?CD pretreatment followed by PPD or PNS interference.The shape of caveolae was changed and the quantity of caveolae was decreased after PPD interference by TEM observation.After CK/Rh2/PPD interference,GM1 was internalized.After PPD interference,Cavl and pCav1 were internalized.After PPD and PNS interference,the mRNA expression of Cav1 was decreased gradually.After PPD treatment,the total protein of Cav1 was decreased at 24h,and the amount of Cav1 in cytoplasm was increased.PPD could activate calcium channel and this activity was down-regulated by M?CD pretreament.The mRNA expression of VEGF was increased at first and then was decreased after PNS interference.The mRNA expression of ICAM-1 was increased at first and then was decreased after PPD and PNS interference.Conclusion The PNS monomer component PPD can stabilize AS plaque by reducing the stability of caveolae/Cav1 to inhibite the proliferation of vascular endothelial cells and exert anti-angiogenic activity. |
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