Profilin acts downstream of LDL to mediate diabetic endothelial cell dysfunction.
Profilin acts downstream of LDL to mediate diabetic endothelial cell dysfunction.
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DOI：
10.1096/fj.03-0841fje
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发表时间：
2004-04-01
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影响因子：
--
通讯作者：
Kazlauskas, Andrius
中科院分区：
文献类型：
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作者：
Romeo, Giulio;Frangioni, John V;Kazlauskas, Andrius
糖尿病患者内皮细胞腔表面发生的变化及其与内皮功能障碍的相关性在体内的特征很差。在这项研究中，我们开发了一种综合策略来发现与糖尿病相关的细胞表面蛋白，并测试它们在内皮功能障碍中的作用。首先，在完整主动脉的内皮表面或糖尿病大鼠和对照大鼠的视网膜内皮细胞中筛选肽噬菌体展示库。然后，我们纯化了profilin-1作为糖尿病相关噬菌体特异性结合伴侣。人糖尿病个体和链脲佐菌素糖尿病大鼠的主动脉内皮中Profilin增加。此外，在大鼠主动脉内皮细胞中过表达profilin触发了内皮功能障碍的3个指标：细胞凋亡增加，ICAM-1表达升高，以及血管舒张剂刺激的磷蛋白（一氧化氮信号传导的标志物）磷酸化降低。ICAM-1和血管舒张刺激磷蛋白的变化在体内糖尿病主动脉中重现。低密度脂蛋白和氧化固醇升高profilin在培养的主动脉内皮细胞。干扰profilin的从头合成消除LDL介导的ICAM-1表达增加。最后，在动脉粥样硬化斑块中，profilin的表达显著升高。这些数据表明，profilin有助于内皮功能障碍的途径，是LDL的下游。
The changes occurring at the luminal surface of endothelial cells in diabetes and their relevance to endothelial dysfunction are poorly characterized in vivo. In this study, we developed an integrated strategy to discover cell surface proteins associated with diabetes and to test their role in endothelial dysfunction. First, a peptide phage display library was screened over the endothelial surface of the intact aorta or in retinal endothelial cells from diabetic and control rats. Then, we purified profilin-1 as a binding partner for one of the diabetic aorta-specific phage. Profilin was increased in the aortic endothelium of human diabetic individuals and streptozotocin-diabetic rats. Furthermore, overexpressing profilin in rat aortic endothelial cells triggered 3 indicators of endothelial dysfunction: increased apoptosis, elevated expression of ICAM-1, and decreased phosphorylation of the vasodilator-stimulated phosphoprotein, a marker for nitric oxide signaling. The changes in ICAM-1 and vasodilator-stimulated phosphoprotein were recapitulated in the diabetic aorta in vivo. LDL and oxysterols elevated profilin in cultured aortic endothelial cells. Interference with the de novo synthesis of profilin abrogated the LDL-mediated increase in ICAM-1 expression. Finally, profilin expression was markedly elevated in atherosclerotic plaques. These data indicate that profilin contributes to endothelial dysfunction in a pathway that is downstream of LDL.