α-Klotho expression determines nitric oxide synthesis in response to FGF-23 in human aortic endothelial cells.
α-Klotho expression determines nitric oxide synthesis in response to FGF-23 in human aortic endothelial cells.
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α-Klotho 表达决定人主动脉内皮细胞响应 FGF-23 的一氧化氮合成
DOI：
10.1371/journal.pone.0176817
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Hsiao LL
中科院分区：
文献类型：
--
作者：
Chung CP;Chang YC;Ding Y;Lim K;Liu Q;Zhu L;Zhang W;Lu TS;Molostvov G;Zehnder D;Hsiao LL
血管内皮细胞表达成纤维细胞生长因子受体，经成纤维细胞生长因子-23处理后具有代谢活性。目前尚不清楚这种作用是α-klotho非依赖性的，还是由体液或内源性内皮细胞α-klotho介导的。在本研究中，我们旨在研究血管内皮细胞α-klotho在人类血管树中的表达，并探讨α-klotho在确定成纤维细胞生长因子-23介导的EC调控中的潜在作用。取人体组织和不同器官的内皮细胞进行免疫组织化学和Western印迹分析。采用人主动脉内皮细胞(HAECs)和人脑微血管内皮细胞(HBMECs)原代培养建立体外细胞模型。除了人脑微血管内皮细胞外，我们发现内源性α-klotho在不同器官的内皮细胞中都有表达。此外，成纤维细胞生长因子-23还可刺激内皮型一氧化氮合酶(ENOS)的表达、一氧化氮(NO)的产生和细胞增殖。有趣的是，在我们的HBMEC体外模型中没有观察到这些影响。高磷处理和内皮细胞α-klotho基因敲除可减轻成纤维细胞生长因子-23介导的eNOS诱导、NO产生和细胞增殖。可溶性α-Klotho的抢救治疗不能逆转HAEC因α-Klotho表达减少或缺失而引起的内皮FGF23抵抗。这些新的观察结果为α-Klotho在人内皮细胞中的不同功能表达提供了证据，它的存在可能在决定血管树对FGF23的反应中发挥作用。在脑微血管内皮细胞中未检测到α-Klotho，其缺失可能使这些细胞对FGF23没有反应。
Endothelial cells (ECs) express fibroblast growth factor (FGF) receptors and are metabolically active after treatment with FGF-23. It is not known if this effect is α-Klotho independent or mediated by humoral or endogenous endothelial α-Klotho. In the present study, we aimed to characterize EC α-Klotho expression within the human vascular tree and to investigate the potential role of α-Klotho in determining FGF-23 mediated EC regulation. Human tissue and ECs from various organs were used for immunohistochemistry and Western blot. Primary cultures of human aortic endothelial cells (HAECs) and human brain microvascular endothelial cells (HBMECs) were used to generate in vitro cell models. We found endogenous α-Klotho expression in ECs from various organs except in microvascular ECs from human brain. Furthermore, FGF-23 stimulated endothelial nitric oxide synthase (eNOS) expression, nitric oxide (NO) production, and cell proliferation in HAECs. Interestingly, these effects were not observed in our HBMEC model in vitro. High phosphate treatment and endothelial α-Klotho knockdown mitigated FGF-23 mediated eNOS induction, NO production, and cell proliferation in HAECs. Rescue treatment with soluble α-Klotho did not reverse endothelial FGF-23 resistance caused by reduced or absent α-Klotho expression in HAECs. These novel observations provide evidence for differential α-Klotho functional expression in the human endothelium and its presence may play a role in determining the response to FGF-23 in the vascular tree. α-Klotho was not detected in cerebral microvascular ECs and its absence may render these cells nonresponsive to FGF-23.