Interventional effect of monomer extract of Radix Salviae Miltiorrhizae on tissue factor gene expression induced by tumor necrosis factor in endothelial cells

Ying SUN; Ziqiang ZHU; Yongmei GAO; Guangjin ZHU

ABSTRACT

BACKGROUND:

It is commonly thought that the expression of tissue factor (TF) gene in human umbilical vein endothelial cells (HUVECs) could be induced by tumor necrosis factor(TNF-α) . But the intervention effect of monomer extract of radix salviae miltiorrhizae(764-3) on TF expression in duced by TNF-α in endothelial cells has not been reported and the mechanism is still unclear.

OBJECTIVE:

To investigate the intervention effect of 764 - 3 on TF expression and calcium ion( [Ca2+] i) induced by TNF-α in HUVECs so as to probe into the possible mechanism of 764 - 3 for preventing cardiovascular thromboembolic disease

DESIGN:

Randomized and controlled trial.

SETTING:

Laboratory of Department of Pathophysiology, Institute of Basic Medical Sciences, Peking Union Medical College MATERIALS This study was conducted in the Department of Pathophysiology, Institute of Basic Medical Science, Peking Union Medical College,Chinese Academy of Medical Sciences from May 1998 to September 1999. Umbilical cord was chosen from Beijing Obstetrics and Gynecology Hospital.

INTERVENTIONS:

ECV304 cell strain and HUVECs were cultured in vitro. With gene recombination techniques, two luciferase reporter genes containing different length of human TF gene promoter were constructed. The two-luciferase reporter genes, together with the intracontrol plasmid pSV-3-gal were respectively cotransfected into cultured ECV304 and HUVECs.MAIN OUTCOME

MEASURES:

The activities of luciferase and βgalactosidase were detected in ECV304 and HUVECs treated by TNF-α or/and 764 -3. Taking Fluo- 3/AM as fluorescent indicator, [Ca2+]i in single HUVEC was observed with laser-scanning confocal microscope.

RESULTS:

The luciferase expression in the p - 244/ + 121 bp luc transfected endothelial cells was significantly increased when the cells were exposed to 100 U/mL TNF-α. The induction of TNF-α could be inhibited by 764 -3 ( P ＜ 0.05). The luciferaseexpression in the p - 111/+ 121 bp Luc transfected endothelial cells was significantly lower than that in the p - 244/+ 121 bp ones and at the same time, 764 -3 did not cause the significantly change of the luciferase expression. Under laser-scanning confocal microscope, TNF-α increased [Ca2 +] i in single HUVEC, but the effect was inhibited by 764 - 3.

CONCLUSION:

TF gene expression induced by TNF-α was inhibited by 764 - 3 in endothelial cells, which was dependent on the p-244/+ 121 bp,and [Ca2+ ]; might be involved in it.