Clivorine, an otonecine pyrrolizidine alkaloid from Ligularia species, impairs neuronal differentiation via NGF-induced signaling pathway in cultured PC12 cells
Clivorine, an otonecine pyrrolizidine alkaloid from Ligularia species, impairs neuronal differentiation via NGF-induced signaling pathway in cultured PC12 cells
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DOI：
10.1016/j.phymed.2016.06.006
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发表时间：
2016-08-15
期刊：
影响因子：
7.9
通讯作者：
Tsim, Karl W. K.
中科院分区：
文献类型：
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作者：
Xiong, Aizhen;Yan, Artemis Lu;Tsim, Karl W. K.
背景：吡咯里西啶生物碱（Pyrrolizidine alkaloids，PAs）广泛存在于多种植物中，包括医药植物。PA的肝毒性已在体内和体外得到证实;然而，PA的神经毒性很少被提及。目的：在这项研究中，我们旨在研究在培养的PC 12细胞中发现的多种Ligularia物种中的PA之一的clivorine的体外神经毒性。研究设计：本研究以PC 12细胞为研究对象，首次阐明了君子兰碱的神经毒性作用及其机制，包括细胞活力和形态学变化，方法：用不同浓度的Clivorine和/或神经生长因子（NGF）刺激PC 12细胞。结果：君子兰碱可抑制细胞增殖，抑制神经元分化，并呈剂量依赖性地抑制神经突起生长。此外，Clivorine降低了编码神经元分化标志物的mRNA的水平，例如神经丝和TrkA（NGF受体）。结论：综上所述，我们的研究结果表明，石蒜碱可能通过下调NGF/TrkA/Akt信号通路而对PC 12细胞产生神经毒性。PAs不仅损害肝脏，而且具有神经毒性，这可能导致大脑疾病，如抑郁症。(C)2016 Elsevier GmbH. All rights reserved.
Background: Pyrrolizidine alkaloids (PAs) are commonly found in many plants including those used in medical therapeutics. The hepatotoxicities of PAs have been demonstrated both in vivo and in vitro; however, the neurotoxicities of PAs are rarely mentioned.Purpose: In this study, we aimed to investigate in vitro neurotoxicities of clivorine, one of the PAs found in various Ligularia species, in cultured PC12 cells.Study design: PC12 cell line was employed to first elucidate the neurotoxicity and the underlying mechanism of clivorine, including cell viability and morphology change, neuronal differentiation marker and signaling pathway.Methods: PC12 cells were challenged with series concentrations of clivorine and/or nerve growth factor (NGF). The cell lysates were collected for MTT assay, trypan blue staining, immunocytofluorescent staining, qRT-PCR and western blotting.Results: Clivorine inhibited cell proliferation and neuronal differentiation evidenced by MTT assay and dose-dependently reducing neurite outgrowth, respectively. In addition, clivorine decreased the level of mRNAs encoding for neuronal differentiation markers, e.g. neurofilaments and TrkA (NGF receptor). Furthermore, clivorine reduced the NGF-induced the phosphorylations of TrkA, protein kinase B and cAMP response element-binding protein in cultured PC12 cells.Conclusion: Taken together, our results suggest that clivorine might possess neurotoxicities in PC12 cells via down-regulating the NGF/TrkA/Akt signaling pathway. PAs not only damage the liver, but also possess neurotoxicities, which could possibly result in brain disorders, such as depression. (C) 2016 Elsevier GmbH. All rights reserved.