| Objective:①To investigate the influence of 17β-Estradiol(E2)on the expression of tissue factor(TF)which is induced by homocysteine (Hcy)in human umbilical vein endothelial cells(HUVEC),and to provide the evidence of Hormone therapy(HT)on vascular endothelial role.②To exploit the principle of NF-βB and estrogen receptor to specifically regulate the E2 on TF expression induced by Hcy in HUVEC.Methods:①Low carbohydrates DMEM was used for culture HUVEC.②Using the method of one-stage clotting assay to measure the total cellular pro-coagulant activity(PCA)of TF.③TF mRNA was measured by semi-quantitative reverse transcription-polymerase chain resction(RT-PCR).④Immunohistochemical analysis was performed to evaluate the variance of NF-κB.Results:①PCA and TF mRNA gradually increased with concentration of Hcy(1×10-5mol/L～1×10-4mol/L)(r=0.969,P＜0.01) in HUVEC,eventually reached the maximum value when Hcy concentration at 1×10-4mol/L.②E2(1×10-9mol/L～1×10-6mol/L) could not inhibit the expression of TF in HUVEC(P＞0.05)by itself directly.③Pretreated with E2(1×10-9mol/L～1×10-6mol/L),it could inhibit the effects of Hcy on PCA and TF mRNA(r=-0.686,P＜0.01),the most effective inhibition concentration level was at 1×10-6mol/L.④ Pure estrogen antagonist ICI182,780 could not significantly inhibit the effects of E2 on TF expression induced by Hey(P＜0.01).⑤Immunohistochemical analysis demonstrated that NF-κB translocation from cytoplasm to the nucleus after treatment of endothelial cells with Hcy(P＜0.01)and the effects of NF-κB translocation induced by Hcy were inhibited by E2(P＜0.01);ICI182,780 could not significantly inhibit the effects of E2 on NF-κB translocation induced by Hcy(P＞0.05).Conclusions:①Hcy could induce the expression of TF in HUVEC. It indicated that Hcy may injury endothelial function.②E2 could not induce the expression of TF in HUVEC alone.It indicated that E2 (1×10-9mol/L～1×10-6mol/L)won't injury endothelial function in nature endothelial cell.②E2 could inhibit the effects of Hcy on the expression of TF,and 1×10-7mol/L was the most effective inhibition concentration.It indicated that E2 may improve endothelial cell injuried endothelial function early.③NF-κB translocation played an important role in Hcy-induced TF expression.E2 could inhibit NF-κB translocation induced by Hcy.④E2 could inhibit the effects of Hcy on the expression of TF,but not through the classic estrogen receptor mediated. |
