Rapamycin regulates the expression and activity of Krüppel-like transcription factor 2 in human umbilical vein endothelial cells.
Rapamycin regulates the expression and activity of Krüppel-like transcription factor 2 in human umbilical vein endothelial cells.
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DOI：
10.1371/journal.pone.0043315
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Zhou Y
中科院分区：
文献类型：
--
作者：
Ma Q;Nie X;Yu M;Wang Z;Yang S;Jia D;Zhou Y
尽管雷帕霉素已被报道可增加人脐静脉内皮细胞（HUVEC）中的促凝血剂和减少抗凝剂，但药物洗脱支架（DES）患者和裸金属支架（BMS）患者的支架血栓形成发生率无显著差异。Krüppel样转录因子2（KLF 2）已被确定为内皮抗血栓功能的关键调节因子。我们假设雷帕霉素可能诱导KLF 2的表达和活性，从而抵消DES诱导的冠状动脉内皮功能障碍。用浓度为2、20、200和2000 ng/ml的雷帕霉素处理HUVECs 24和48小时，不加或加凝血酶，评估KLF 2、组织因子（TF）和内皮型一氧化氮合酶（eNOS）的表达。雷帕霉素高剂量组KLF 2表达和活性明显增强（p<0.01）。与对照组相比，雷帕霉素组TF表达增加，24 h后抑制eNOS表达（p<0.01）。此外，小干扰RNA介导的KLF 2敲低强烈放大了雷帕霉素诱导TF的能力，并减少内皮细胞中eNOS的积累。雷帕霉素依赖性诱导KLF 2表达可能部分抵消冠状动脉内皮功能障碍，从而为预防DES诱导的支架内血栓形成提供了一个新的分子靶点。
Although rapamycin has been reported to increase procoagulants and decrease anticoagulants in human umbilical vein endothelial cells (HUVECs), there is no significant difference in the incidence of stent thrombosis between patients with drug-eluting stents (DESs) and those with bare metal stents (BMSs). Krüppel-like transcription factor 2 (KLF2) has been identified as a key regulator of endothelial antithrombotic function. We hypothesized that rapamycin might induce the expression and activity of KLF2, thereby counteracting coronary endothelial dysfunction induced by DESs. Expression of KLF2, tissue factor (TF) and endothelial NO synthase (eNOS) were assessed in HUVECs treated with rapamycin at concentrations of 2, 20, 200 and 2000 ng/ml for 24 and 48 hours without or with thrombin. Rapamycin strongly induced the expression and activity of KLF2 in high dose groups (p<0.01). Compared with control group, the expression of TF was increased by rapamycin, which inhibited the expression of eNOS after treating for 24 hours (p<0.01). Furthermore, small-interfering RNA–mediated knockdown of KLF2 strongly magnified the ability of rapamycin to induce TF and reduce eNOS accumulation in HUVECs. Rapamycin-dependent induction of KLF2 might partly counteract coronary endothelial dysfunction and thereby provided a novel molecular target to prevent stent thrombosis induced by DESs.