丹参对血管紧张素Ⅱ诱导内皮细胞组织因子基因表达的作用

Effect of Danshen on TF expression induced by angiotensin Ⅱ in endothelial cells

目的 探讨丹参有效单体764-3(764-3)对心血管血栓栓塞性疾病防治的可能机制。方法 体外培养人脐静脉内皮细胞(HUVEC)、内皮细胞株(ECV304)。采用限制性内切酶法,构建含有人血管内皮细胞组织因子基因不同上游调控序列的荧光素酶报告基因质粒,经脂质体法转染内皮细胞,用血管紧张素Ⅱ(AngⅡ)及764-3处理内皮细胞,测定细胞裂解物荧光素酶及β半乳糖苷酶活性。用激光共聚焦扫描显像系统观测单个内皮细胞胞内游离钙浓度([Ca2+]i)水平的改变。结果 在TF基因上游序列-244/+121bp存在下,AngⅡ可使转染内皮细胞荧光素酶表达量明显高于对照组,764-3抑制这种增加。AngⅡ可引起人内皮细胞[Ca2+]i缓慢升高,加入764-3可迅速大幅度降低[Ca2+]i,使接近基线水平。结论764-3可抑制AngⅡ诱导的人内皮细胞TF基因表达增强,这种作用依赖于该基因上游序列-244/+121bp的存在。

Objective To investigate the intervenient effect of effective monoploid of Danshen 764-3 on TF( tissue factor) expression and calcium ion([Ca2+ ]i) induced by angiotensin Ⅱ(AngⅡ ) in human vascular endo-thelial cells. Methods Two luciferase reporter genes containing different upstream sequences of TF gene were constructed. The two luciferase reporter genes together with the intracontrol plasmid pSV-β-gal were co-transfected respectively into cultured human vascular endothelial cells(HUVEC) .The relative luciferase activities were detected in endothelial cells ECV304 treated by Ang][ or/and 764-3. [Ca2+ ]i in single HUVEC was observed with laser-scanning confocal microscope. Results A ng Ⅱ could significantly increase the luciferase expression in the p-244/ + 121bp Luc transfected endothelial cells and the effect of AngⅡ could be inhibited by 764-3. AngⅡ could increase [Ca2+ ]i in single HUVEC, but it could be inhibited by 764-3. Conclusion TF gene expression induced by AngⅡ could be inhibited by 764-3 and [Ca2+ ]i was involved in it.