Neuroprotection Research Laboratory, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, Mass., USA.
Cerebrovasc Dis. 2010 Aug;30(3):290-6. doi: 10.1159/000319072. Epub 2010 Jul 23.
Increasing evidence suggests that beyond its antiplatelet properties, dipyridamole may have pleiotropic effects on other cells within the neurovascular elements of the brain. In this experimental cellular study, we asked whether dipyridamole can ameliorate brain endothelial injury after exposure to inflammatory and metabolic insults.
Human brain endothelial cells were grown in culture, and exposed to TNFalpha (continuously for 20 h) or subjected to oxygen-glucose deprivation (OGD; 6 h of insult followed by 18 h recovery). Expression of ICAM-1, VCAM-1 and PECAM-1 were measured by immunoblotting. Matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in the conditioned media were quantified via zymography. MTT mitochondrial activity was measured to assess endothelial cell viability.
Exposure of human brain endothelial cells to TNFalpha (12.5-50 ng/ml) induced a clear increase in protein levels of ICAM-1, VCAM-1 and MMP-9. TNFalpha did not alter PECAM-1. Dipyridamole (1-5 muM) significantly attenuated ICAM-1 and MMP-9 levels after this inflammatory insult. No significant effects of dipyridamole were noted for VCAM-1. Six-hour OGD induced moderate endothelial cell death accompanied by a release of MMP-9. Dipyridamole significantly decreased MMP-9 levels and cell death after this metabolic insult.
These results suggest that dipyridamole may ameliorate brain endothelial injury after inflammation and/or metabolic insults. How these putative cellular mechanisms may relate to clinical outcomes and conditions in stroke patients remains to be elucidated.
越来越多的证据表明，双嘧达莫除了具有抗血小板作用外，还可能对大脑神经血管单元中的其他细胞产生多效性影响。在这项实验性细胞研究中，我们想知道双嘧达莫是否可以减轻暴露于炎症和代谢损伤后脑内皮细胞的损伤。
将人脑内皮细胞在培养中生长，并暴露于 TNFα（连续 20 小时）或进行氧葡萄糖剥夺（6 小时的损伤，然后 18 小时恢复）。通过免疫印迹法测量 ICAM-1、VCAM-1 和 PECAM-1 的表达。通过酶谱法定量测定条件培养基中的基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）。通过 MTT 线粒体活性测定评估内皮细胞活力。
TNFα（12.5-50ng/ml）暴露于人脑内皮细胞可明显增加 ICAM-1、VCAM-1 和 MMP-9 的蛋白水平。TNFα不改变 PECAM-1。双嘧达莫（1-5μM）在这种炎症损伤后明显降低 ICAM-1 和 MMP-9 水平。双嘧达莫对 VCAM-1 没有明显影响。6 小时 OGD 诱导内皮细胞中度死亡，并伴有 MMP-9 的释放。双嘧达莫可显著降低代谢损伤后 MMP-9 水平和细胞死亡。
这些结果表明，双嘧达莫可能减轻炎症和/或代谢损伤后的脑内皮损伤。这些潜在的细胞机制如何与中风患者的临床结果和情况相关仍有待阐明。
