Chronic administration of BMS309403 improves endothelial function in apolipoprotein E-deficient mice and in cultured human endothelial cells
Chronic administration of BMS309403 improves endothelial function in apolipoprotein E-deficient mice and in cultured human endothelial cells
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DOI：
10.1111/j.1476-5381.2010.01158.x
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发表时间：
2011-04-01
影响因子：
7.3
通讯作者：
Vanhoutte, Paul M.
中科院分区：
文献类型：
--
作者：
Lee, Mary Y. K.;Li, Huiying;Vanhoutte, Paul M.
背景与目的脂肪细胞脂肪酸结合蛋白（A-FABP）在再生内皮细胞中上调，并调节巨噬细胞的炎症反应。伴随再生的内皮功能障碍可因高脂血症而加速。在这里，我们研究了A-FABP在载脂蛋白e缺乏（ApoE-/-）小鼠和培养的人内皮细胞主动脉内皮功能障碍的发病机制中的作用。实验方法采用RT-PCR、免疫染色和免疫印迹法检测ApoE-/-小鼠主动脉和人内皮细胞中ha - fabp的含量。用免疫印迹法测定内皮型一氧化氮合酶（eNOS）的总形式和磷酸化形式。结果12周龄及以上ApoE-/-小鼠主动脉内皮中表达sa - fabp， 8周龄及C57野生型小鼠主动脉内皮中无表达。与年龄匹配的对照组相比，18周龄ApoE-/-小鼠主动脉中乙酰胆碱、UK14304（选择性α(2)-肾上腺素能受体激动剂）和A23187（钙离子载体）的内皮依赖性松弛减少，磷酸化eNOS和总eNOS的蛋白含量减少。在12周龄小鼠中，A- fabp抑制剂BMS309403治疗了6周，改善了内皮功能，磷酸化和总eNOS，降低了血浆甘油三酯水平，但不影响内皮非依赖性松弛。BMS309403对uk14304所致松弛的有益作用被百日咳毒素减弱。在培养的人微血管内皮细胞中，脂质诱导的A-FABP表达与磷酸化eNOS和NO产生的减少有关，BMS309403可以逆转这一现象。结论和意义内皮细胞中A-FABP的表达升高有助于其体内和体外功能障碍。
BACKGROUND AND PURPOSEAdipocyte fatty acid-binding protein (A-FABP) is up-regulated in regenerated endothelial cells and modulates inflammatory responses in macrophages. Endothelial dysfunction accompanying regeneration is accelerated by hyperlipidaemia. Here, we investigate the contribution of A-FABP to the pathogenesis of endothelial dysfunction in the aorta of apolipoprotein E-deficient (ApoE-/-) mice and in cultured human endothelial cells.EXPERIMENTAL APPROACHA-FABP was measured in aortae of ApoE-/-mice and human endothelial cells by RT-PCR, immunostaining and immunoblotting. Total and phosphorylated forms of endothelial nitric oxide synthase (eNOS) were measured by immunoblotting. Changes in isometric tension were measured in rings of mice aortaeKEY RESULTSA-FABP was expressed in aortic endothelium of ApoE-/- mice aged 12 weeks and older, but not at 8 weeks or in C57 wild-type mice. Reduced endothelium-dependent relaxations to acetylcholine, UK14304 (selective alpha(2)-adrenoceptor agonist) and A23187 (calcium ionophore) and decreased protein presence of phosphorylated and total eNOS were observed in aortae of 18 week-old ApoE-/- mice compared with age-matched controls. A 6 week treatment with the A-FABP inhibitor, BMS309403, started in 12 week-old mice, improved endothelial function, phosphorylated and total eNOS and reduced plasma triglyceride levels but did not affect endothelium-independent relaxations. The beneficial effect of BMS309403 on UK14304-induced relaxations was attenuated by Pertussis toxin. In cultured human microvascular endothelial cells, lipid-induced A-FABP expression was associated with reduced phosphorylated eNOS and NO production and was reversed by BMS309403.CONCLUSIONS AND IMPLICATIONSElevated expression of A-FABP in endothelial cells contributes to their dysfunction both in vivo and in vitro.