Department of Physiology, Defence Institute of Physiology and Allied Sciences, Timarpur, New Delhi, India.
Department of Physiology, HIMSR, Jamia Hamdard, Hamdard Nagar, New Delhi, India.
J Leukoc Biol. 2019 Apr;105(4):705-717. doi: 10.1002/JLB.2A0918-361R. Epub 2019 Mar 5.
PGE plays a critical role in angiogenesis, ischemic, and neuro-inflammatory disorders of the brain, which breakdown the blood-brain barrier (BBB). However, the effects of PGE on human brain endothelial cell (HBECs) migration, a key process in the angiogenic response and BBB stability, are not well defined. In this study, we investigated the mechanism of PGE in HBECs migration in vitro. Here we showed that PGE stimulated migration of HBECs in a dose-time and matrix-dependent manner, evaluated by the Boyden chamber assay, but other prostanoids failed to do so. PGE receptor (EP2; butaprost), EP3 (sulprostone), and EP4 (PGE -OH) receptor agonists stimulated HBECs migration, but the silencing of EP significantly attenuated this effect. EP1 agonist (11-trinor PGE ) had no effect on HBECs migration on silencing of the EP1 receptor. We further showed that PGE stimulated cAMP production and activated protein kinase A (PKA), whereas pretreatment with the adenyl cyclase inhibitor (dideoxyadenosine; 1 μM) or PKA inhibitors, H89 (0.5 μM)/PKAI (1 μM), completely abrogated PGE -induced migration. Furthermore, silencing of the EP2/EP4 receptors significantly inhibited PGE -induced cAMP and PKA activation, whereas EP3 receptor silencing failed to do so. These results suggest that PGE regulates HBEC migration via cooperation of EP2, EP3, and EP4 receptors. Coupling of PGE to these receptors resulted in increased production of cAMP, which regulates HBEC migration via PKA pathway. The elucidation of molecular events involved is critical for the development of targeted strategies to treat cerebrovascular diseases associated with dysregulated angiogenesis.
前列腺素 E（PGE）在脑的血管生成、缺血和神经炎症紊乱中起着关键作用，这些紊乱会破坏血脑屏障（BBB）。然而，PGE 对人脑血管内皮细胞（HBECs）迁移的影响，即血管生成反应和 BBB 稳定性的关键过程，尚未得到很好的定义。在这项研究中，我们研究了 PGE 在体外刺激 HBECs 迁移的机制。我们发现 PGE 以剂量和基质依赖的方式刺激 HBECs 的迁移，通过 Boyden 室测定评估，但其他前列腺素则不能。PGE 受体（EP2；butaprost）、EP3（sulprostone）和 EP4（PGE-OH）受体激动剂刺激 HBECs 迁移，但 EP 的沉默显著减弱了这种作用。EP1 激动剂（11-trinor PGE）在 EP1 受体沉默时对 HBECs 迁移没有影响。我们进一步表明，PGE 刺激 cAMP 的产生并激活蛋白激酶 A（PKA），而用腺苷酸环化酶抑制剂（二脱氧腺苷；1 μM）或 PKA 抑制剂 H89（0.5 μM）/PKAI（1 μM）预处理完全阻断了 PGE 诱导的迁移。此外，EP2/EP4 受体的沉默显著抑制了 PGE 诱导的 cAMP 和 PKA 激活，而 EP3 受体的沉默则不能。这些结果表明，PGE 通过 EP2、EP3 和 EP4 受体调节 HBEC 迁移。PGE 与这些受体的偶联导致 cAMP 的产生增加，cAMP 通过 PKA 途径调节 HBEC 迁移。阐明所涉及的分子事件对于开发针对与血管生成失调相关的脑血管疾病的靶向策略至关重要。
