首页> 外文期刊>Journal of Neurochemistry: Offical Journal of the International Society for Neurochemistry >Caveolin-1 mediates tissue plasminogen activator-induced MMP-9 up-regulation in cultured brain microvascular endothelial cells
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Caveolin-1 mediates tissue plasminogen activator-induced MMP-9 up-regulation in cultured brain microvascular endothelial cells

机译:Caveolin-1在培养的脑微血管内皮细胞中介导组织纤溶酶原激活物诱导的MMP-9上调。

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摘要

Thrombolysis with tissue plasminogen activator (tPA) increases matrix metalloproteinase-9 (MMP-9) activity in the ischemic brain, which exacerbates blood-brain barrier injury and increases the risk of symptomatic cerebral hemorrhage. The mechanism through which tPA enhances MMP-9 activity is not well understood. Here we report an important role of caveolin-1 in mediating tPA-induced MMP-9 synthesis. Brain microvascular endothelial cell line bEnd3 cells were incubated with 5 or 20 lg/ml tPA for 24 hrs before analyzing MMP-9 levels in the conditioned media and cellular extracts by gelatin zymography. tPA at a dose of 20 lg/mL tPA, but not 5 lg/mL, significantly increased MMP-9 level in cultured media while decreasing it in cellular extracts. Concurrently, tPA treatment induced a 2.3-fold increase of caveolin-1 protein levels in endothelial cells. Interestingly, knockdown of Cav-1 with siRNA inhibited tPAinduced MMP-9 mRNA up-regulation and MMP-9 increase in the conditioned media, but did not affect MMP-9 decrease in cellular extracts. These results suggest that caveolin-1 critically contributes to tPA-mediated MMP-9 up-regulation, but may not facilitate MMP-9 secretion in endothelial cells.
机译:用组织纤溶酶原激活剂(tPA)进行溶栓可增加缺血性脑中的基质金属蛋白酶9(MMP-9)活性,从而加剧血脑屏障损伤并增加症状性脑出血的风险。 tPA增强MMP-9活性的机制尚不清楚。在这里,我们报告小窝蛋白1在介导tPA诱导的MMP-9合成中的重要作用。将脑微血管内皮细胞系bEnd3细胞与5或20 lg / ml tPA孵育24小时,然后通过明胶酶谱分析条件培养基和细胞提取物中的MMP-9水平。 tPA剂量为20 lg / mL tPA而不是5 lg / mL时,显着提高了培养基中的MMP-9水平,同时降低了细胞提取物中的MMP-9水平。同时,tPA处理可诱导内皮细胞小窝蛋白1蛋白水平增加2.3倍。有趣的是,用siRNA敲低Cav-1可抑制tPA诱导的MMP-9 mRNA上调,并在条件培养基中增加MMP-9,但不影响细胞提取物中MMP-9的减少。这些结果表明小窝蛋白1关键有助于tPA介导的MMP-9上调,但可能不促进内皮细胞中MMP-9的分泌。

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