Luteolin stimulates the NGF-induced neurite outgrowth in cultured PC12 cells through binding with NGF and potentiating its receptor signaling
Luteolin stimulates the NGF-induced neurite outgrowth in cultured PC12 cells through binding with NGF and potentiating its receptor signaling
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DOI：
10.1039/d1fo01096d
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发表时间：
2021-10-05
期刊：
影响因子：
6.1
通讯作者：
Tsim, Karl Wah-Keung
中科院分区：
文献类型：
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作者：
Gao, Alex Xiong;Xia, Tracy Chen-Xi;Tsim, Karl Wah-Keung
木犀草素是水果和蔬菜中的一种类黄酮，具有神经营养功能，但其机制尚不清楚。在这里，我们假设木犀草素与神经生长因子 (NGF) 存在直接相互作用；因此，NGF 的功能可以得到增强。通过超滤、Biacore 和对接分析验证了木犀草素与 NGF 的直接结合。在培养的 PC12 细胞中，木犀草素与低剂量 NGF 联合应用可增强 NGF 诱导的神经元分化，使分化细胞数量增加至 25.4 +/- 4.8% (p < 0.01)，并使神经丝表达增加 119 +/- 32.1% (p < 0.05)、191 +/- 12.6% (p < 0.05)。 NF68、NF160 和 NF200 分别为 110 +/- 23.4% (p < 0.05) 和 110 +/- 23.4% (p < 0.05)。联合治疗诱导原肌球蛋白受体激酶 A (TrkA)、细胞外信号调节激酶 1/2 (ERK1/2)、蛋白激酶 B (Akt)、磷脂酶 C-gamma 1 (PLC gamma 1) 和 cAMP 反应元件结合蛋白 (CREB) 磷酸化 2 至 3 倍：这些诱导的磷酸化模仿高剂量 NGF 的磷酸化。此外，TrkA 抑制剂 K252a 的应用阻断了培养的 PC12 细胞中木犀草素介导的神经丝表达和神经突生长的诱导，表明了目标特异性。该结果支持开发木犀草素作为 NGF 缺乏相关神经退行性疾病的治疗或预防剂。
Luteolin, a flavonoid in fruits and vegetables, has neurotrophic functions without a well-characterized mechanism. Here, we hypothesize a direct interaction of luteolin with nerve growth factor (NGF); as such, the functionality of the NGF could be potentiated. The direct binding of luteolin with NGF was validated by ultra-filtration, Biacore, and docking analyses. In cultured PC12 cells, application of luteolin in combination with a low dose of NGF potentiated the NGF-induced differentiation of neurons by an increase of the differentiated cell number to 25.4 +/- 4.8% (p < 0.01), as well as the increased expression of neurofilaments by 119 +/- 32.1% (p < 0.05), 191 +/- 12.6% (p < 0.01), and 110 +/- 23.4% (p < 0.05) for NF68, NF160 and NF200, respectively. The co-treatment induced the phosphorylations of tropomyosin receptor kinase A (TrkA), extracellular signal-regulated kinase 1/2 (ERK1/2), protein kinase B (Akt), phospholipase C-gamma 1 (PLC gamma 1), and cAMP response element-binding protein (CREB) by 2 to 3 fold: these induced phosphorylations were mimicking that of a high dose of NGF. Moreover, the application of the TrkA inhibitor, K252a, blocked the luteolin-mediated induction of neurofilament expression and neurite outgrowth in cultured PC12 cells, suggesting the target specificity. The result supports the development of luteolin as a therapeutic, or preventive, agent for NGF insufficiency-associated neurodegenerative diseases.