Effect of curcumin on the adhesion of platelets to brain microvascular endothelial cells in vitro
Abstract
Aim: To determine whether curcumin prevents the adhesion of platelets to brain microvascular endothelial cells (BMECs) cultured in vitro.
Methods: [3H]Adenine-labeled platelets were incubated with BMECs to investigate the role of curcumin in the adhesion of platelets to BMECs. The number of platelets adhering to the BMECs monolayer was determined by liquid scintillation spectroscopy. The thrombin-induced expression of platelets P-selectin, glycoprotein IIb (GPIIb), and glycoprotein IIIa (GPIIIa) on the cell surface, was measured by flow cytometry. P-selectin mRNA levels of BMECs were determined by RT-PCR. The TNF-α-induced expressions of P-selectin and E-selectin on the surface of BMECs were determined by Western blotting.
Results: The adhesion between thrombin-activated platelets and normal BMECs, and that of TNF-α-activated BMECs and normal platelets were significantly increased, and this increase could be inhibited by curcumin (30–240 μmol/L) in a concentration-dependant manner. The platelets activated with thrombin and BMECs stimulated by TNF-α demonstrated an upregulated expressions of P-selectin and E-selectin, and this increase, when pretreated with curcumin for 30 min, could be restrained dose dependently. Curcumin also inhibited the increase of the GPIIb/GPIIIa expression of thrombin-activated platelets in a concentration-dependent manner.
Conclusion: Curcumin can inhibit the platelets to BMECs. This effect may be related to the decreased expressions of P-selectin, E-selectin, and GPIIb/GPIIIa on platelets and BMECs.
Keywords:
Methods: [3H]Adenine-labeled platelets were incubated with BMECs to investigate the role of curcumin in the adhesion of platelets to BMECs. The number of platelets adhering to the BMECs monolayer was determined by liquid scintillation spectroscopy. The thrombin-induced expression of platelets P-selectin, glycoprotein IIb (GPIIb), and glycoprotein IIIa (GPIIIa) on the cell surface, was measured by flow cytometry. P-selectin mRNA levels of BMECs were determined by RT-PCR. The TNF-α-induced expressions of P-selectin and E-selectin on the surface of BMECs were determined by Western blotting.
Results: The adhesion between thrombin-activated platelets and normal BMECs, and that of TNF-α-activated BMECs and normal platelets were significantly increased, and this increase could be inhibited by curcumin (30–240 μmol/L) in a concentration-dependant manner. The platelets activated with thrombin and BMECs stimulated by TNF-α demonstrated an upregulated expressions of P-selectin and E-selectin, and this increase, when pretreated with curcumin for 30 min, could be restrained dose dependently. Curcumin also inhibited the increase of the GPIIb/GPIIIa expression of thrombin-activated platelets in a concentration-dependent manner.
Conclusion: Curcumin can inhibit the platelets to BMECs. This effect may be related to the decreased expressions of P-selectin, E-selectin, and GPIIb/GPIIIa on platelets and BMECs.