Zhang Li, Gu Zhen-lun, Qin Zheng-hong, Liang Zhong-qin
Department of Pharmacology and Laboratory of Aging and Nervous Diseases, Soochow University School of Medicine, Suzhou 215123, China.
Acta Pharmacol Sin. 2008 Jul;29(7):800-7. doi: 10.1111/j.1745-7254.2008.00813.x.
To determine whether curcumin prevents the adhesion of platelets to brain microvascular endothelial cells (BMECs) cultured in vitro.
[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-alpha- induced expressions of P-selectin and E-selectin on the surface of BMECs were determined by Western blotting.
The adhesion between thrombin-activated platelets and normal BMECs, and that of TNF-alpha-activated BMECs and normal platelets were significantly increased, and this increase could be inhibited by curcumin (30-240 micromol/L) in a concentration-dependant manner. The platelets activated with thrombin and BMECs stimulated by TNF-alpha 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 thrombinactivated platelets in a concentration-dependent manner.
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.
确定姜黄素是否能预防体外培养的血小板与脑微血管内皮细胞(BMECs)的黏附。
将[³H]腺嘌呤标记的血小板与BMECs共同孵育,以研究姜黄素在血小板与BMECs黏附中的作用。通过液体闪烁光谱法测定黏附于BMECs单层的血小板数量。采用流式细胞术检测凝血酶诱导的血小板表面P-选择素、糖蛋白IIb(GPIIb)和糖蛋白IIIa(GPIIIa)的表达。通过逆转录聚合酶链反应(RT-PCR)测定BMECs的P-选择素mRNA水平。采用蛋白质免疫印迹法检测肿瘤坏死因子-α(TNF-α)诱导的BMECs表面P-选择素和E-选择素的表达。
凝血酶激活的血小板与正常BMECs之间以及TNF-α激活的BMECs与正常血小板之间的黏附显著增加,而姜黄素(30 - 240微摩尔/升)可以浓度依赖性地抑制这种增加。凝血酶激活的血小板和TNF-α刺激的BMECs表现出P-选择素和E-选择素表达上调,而用姜黄素预处理30分钟后,这种增加可呈剂量依赖性受到抑制。姜黄素还以浓度依赖性方式抑制凝血酶激活的血小板中GPIIb/GPIIIa表达的增加。
姜黄素可抑制血小板与BMECs的黏附。这种作用可能与血小板和BMECs上P-选择素、E-选择素以及GPIIb/GPIIIa表达降低有关。