The Study Of Protective Effects Of Tetramethylpyrazine(TMP),Astragalus Polysaccharides (APS) And Their Combination On Vascular Endothelial Cells(VECs)

ObjectiveBy establishing human umbilical vein endothelial cells lines(ECV-304)injury model associated with serum of patients with hypertension disease,to observe the effct of ligustrazine (TMP),astragalus polysaccharides(APS)and monomer combination on activeness,shape,secretion function and cells'intracellular free calcium([Ca²⁺]i)of the modle cells,to research the protective effects of the tradition chinese medicine(TCM)on vascular endothelial cells(VECs).MethodsTo establish the cell modle,We select that the secum concentrations is 10%and the time is 24h,at the same time ECV-304 were processed by TMP,APS and their different combinations. Esperiments were divided into 5 groups:the modle group,the control group,the TMP groups,the APS groups,the monomer combination groups.the activeness of cultivated ECV-304 were assayed by MTT chromatometry,the change of the cells'shape was observed by the inverse phase contrast microscope and HE staining,the nitric oxide(NO)excreted by the cells was assayed by nitric acid deoxidizing enzyme method,and the endothelin(ET)was assayed by non-balance method,the cells'intracellular free calcium([Ca²⁺]i)was assayed by laser scanning confocal microscope(LSCM).Results1.Cells'activeness:After the serum acted on cells for 24h,the cells'activeness was lower in the modle group than in the control group;When the TMP concentration is 160μg/ml,80μg/ml,40μg/ml,the APS concentration is 640μg/ml,160μg/ml,80μg/ml,the cells'activeness was higher in the medicine group than in the modle group;the cells'activeness was more higher in the TMP combinate with APS groups than in the a single medicine groups.2.Cells'shape:By the inverse phase contrast microscope and HE staining,we discovered that the difference of shape was distinct between the modle group and the control group,the medicine can alleviated the morphological damage partly with medicine concentrations increasing.3.Cells'secretion function:Model group decreased the NO level and increased ET level significantly by comparison of control group(P＜0.01),A single medicine groups and combinated groups increased the NO level and decreased ET level significantly compared with the model group(P＜0.01),but at the low dosage group medicine had no obvious effects;The content of NO and ET excreted by cells the difference between the TMP and APS combinated group and a single medicine group was distinct(P＜0.01).4.Cells'intracellular free calcium:Fluorescence intensity was high in the model group than in the control group,the difference was distinct(P＜0.01).Fluorescence intensity was lower in the drug group than in the model group,the difference was distinct(P＜0.01 or P＜0.05).Compared the TMP and APS combinated group with a single medicine group, Fluorescence intensity was decreased,the difference was distinct(P＜0.01).ConclusionWe think that TMP and ASP has the distinct protective effects on VECs.We also found that TMP combinated with APS is superior to a single medicine on the protective effects on VECs.