Fluid shear stress upregulates placental growth factor in the vessel wall via NADPH oxidase 4.
Fluid shear stress upregulates placental growth factor in the vessel wall via NADPH oxidase 4.
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DOI：
10.1152/ajpheart.00408.2015
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发表时间：
2015-11
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影响因子：
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通讯作者：
N. Rashdan;P. Lloyd
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作者：
N. Rashdan;P. Lloyd
胎盘生长因子（PLGF）是一种有效的动脉生成刺激物，在动脉重构过程中被上调。流体剪切应力（FSS）的增加是动脉发生的关键生理刺激。然而，FSS在调节PLGF表达中的作用尚不清楚。为了验证FSS调节PLGF在血管细胞中的表达的假设，并确定其参与的信号通路，我们在多孔Transwell插入物的两侧培养人冠状动脉内皮细胞（HCAEC）和人冠状动脉平滑肌细胞。然后将HCAEC暴露于0.07 Pa（“正常”，模拟静止侧枝的流动）、1.24 Pa（“高”，模拟重塑侧枝的流动）或0.00 Pa（“静态”）的脉动FSS中2小时。与静态对照相比，高FSS使分泌的PLGF蛋白增加了1.4倍（n = 5, P < 0.01），而正常FSS对PLGF没有显著影响。同样，高流量刺激离体小鼠肠系膜小动脉PLGF mRNA表达几乎翻倍。使用siRNA敲除PLGF发现HCAEC是共培养PLGF的主要来源（n = 5, P < 0.01）。与静态对照相比，FSS可提高H2O2和一氧化氮的产量（n = 5, P < 0.05）。N(G)-硝基精氨酸甲酯（100 μM）对fss诱导的PLGF升高无显著影响。过氧化氢酶（500 U/ml）和二苯二酮（5 μM）均能减弱FSS对共培养PLGF蛋白的影响。二苯乙酮也阻断了高流量对离体小动脉中PLGF mRNA上调的作用。进一步的研究发现NADPH氧化酶4是该途径的活性氧来源。我们得出结论，FSS通过NADPH氧化酶4和活性氧信号调节PLGF的表达。
Placental growth factor (PLGF), a potent stimulator of arteriogenesis, is upregulated during outward arterial remodeling. Increased fluid shear stress (FSS) is a key physiological stimulus for arteriogenesis. However, the role of FSS in regulating PLGF expression is unknown. To test the hypothesis that FSS regulates PLGF expression in vascular cells and to identify the signaling pathways involved, human coronary artery endothelial cells (HCAEC) and human coronary artery smooth muscle cells were cultured on either side of porous Transwell inserts. HCAEC were then exposed to pulsatile FSS of 0.07 Pa ("normal," mimicking flow through quiescent collaterals), 1.24 Pa ("high," mimicking increased flow in remodeling collaterals), or 0.00 Pa ("static") for 2 h. High FSS increased secreted PLGF protein ∼1.4-fold compared with static control (n = 5, P < 0.01), while normal FSS had no significant effect on PLGF. Similarly, high flow stimulated PLGF mRNA expression nearly twofold in isolated mouse mesenteric arterioles. PLGF knockdown using siRNA revealed that HCAEC were the primary source of PLGF in cocultures (n = 5, P < 0.01). Both H2O2 and nitric oxide production were increased by FSS compared with static control (n = 5, P < 0.05). N(G)-nitro-l-arginine methyl ester (100 μM) had no significant effect on the FSS-induced increase in PLGF. In contrast, both catalase (500 U/ml) and diphenyleneiodonium (5 μM) attenuated the effects of FSS on PLGF protein in cocultures. Diphenyleneiodonium also blocked the effect of high flow to upregulate PLGF mRNA in isolated arterioles. Further studies identified NADPH oxidase 4 as a source of reactive oxygen species for this pathway. We conclude that FSS regulates PLGF expression via NADPH oxidase 4 and reactive oxygen species signaling.