Combination of Reactive Oxygen Species and Tissue-Type Plasminogen Activator Enhances the Induction of Gelatinase B in Brain Endothelial Cells
Combination of Reactive Oxygen Species and Tissue-Type Plasminogen Activator Enhances the Induction of Gelatinase B in Brain Endothelial Cells
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DOI：
10.3109/00207454.2011.623808
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发表时间：
2012-02-01
影响因子：
2.2
通讯作者：
Umemura, Kazuo
中科院分区：
文献类型：
--
作者：
Harada, Kosuke;Suzuki, Yasuhiro;Umemura, Kazuo
组织型纤溶酶原激活剂（t-PA）通过脑内皮细胞中明胶酶B（基质金属蛋白酶-9; MMP-9）的产生增加颅内出血的风险。最近报道，自由基清除剂依达拉奉显著降低t-PA介导的MMP-9的产生。因此，通过使用培养的脑内皮细胞（b.End3），我们调查是否t-PA介导的MMP-9的生产增强活性氧（ROS）及其信号通路。此外，我们还研究了依达拉奉是否能降低这种联合增强。将b.End3细胞暴露于t-PA（10 μ g/mL），然后暴露于H2 O2（30 μ M）;进一步，测量MMP-9蛋白水平。ROS促进MMP-9的产生，并且ROS + t-PA比单独的t-PA或ROS更显著地增加MMP-9的产生。结果表明，单独使用H_2O_2或t-PA可显著增加NF-κ B向细胞核的转运，而t-PA和H_2O_2联合使用可更大程度地增加NF-κ B向细胞核的转运。此外，t-PA和ROS的组合显著增加I-κ B降解以及NF-κ B表达。依达拉奉完全降低ROS加t-PA介导的MMP-9增强。总之，ROS增强了脑内皮细胞中t-PA介导的MMP-9的产生;加入依达拉奉可降低MMP-9的产生，依达拉奉可抑制NF-κ B通路，特别是通过增强I-κ B降解。
Tissue-type plasminogen activator (t-PA) increases the risk of intracranial hemorrhage by gelatinase B (matrix metalloprotease-9; MMP-9) production in brain endothelial cells. It was recently reported that the free-radical scavenger edaravone significantly decreases t-PA-mediated MMP-9 production. Therefore, by using cultured brain endothelial cells (b.End3), we investigated whether t-PA-mediated MMP-9 production was enhanced by reactive oxygen species (ROS) and its signaling pathways. Moreover, we also investigated that whether this combined enhancement is reduced by edaravone. The b.End3 cells were exposed to t-PA (10 mu g/mL), followed by H2O2 (30 mu M); further, the MMP-9 protein level was measured. ROS enhanced MMP-9 production, and ROS plus t-PA significantly increased MMP-9 production more than t-PA or ROS alone. The results showed that H2O2 or t-PA alone caused a significant increase in NF-kappa B translocation to the nucleus, whereas the combination of t-PA and H2O2 increased the translocation of NF-kappa B to an even greater extent. Moreover, the combination of t-PA and ROS significantly increased I-kappa B degradation as well as NF-kappa B expression. Edaravone completely decreased the ROS plus t-PA-mediated MMP-9 enhancement. In conclusion, ROS enhanced t-PA-mediated MMP-9 production in brain endothelial cells; this MMP-9 production was decreased by the addition of edaravone, which inhibited the NF-kappa B pathway, specifically by enhancing I-kappa B degradation.