Paeoniflorin, a potent natural compound, protects PC12 cells from MPP+ and acidic damage via autophagic pathway
Paeoniflorin, a potent natural compound, protects PC12 cells from MPP+ and acidic damage via autophagic pathway
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DOI：
10.1016/j.jep.2010.06.009
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发表时间：
2010-08-19
影响因子：
5.4
通讯作者：
Liu, Chun-Feng
中科院分区：
文献类型：
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作者：
Cao, Bi-Yin;Yang, Ya-Ping;Liu, Chun-Feng
民族药理学相关性：芍药苷（Paeoniflorin， PF）是白芍的主要生物活性成分，在中药中被广泛用于治疗神经退行性疾病，如帕金森病（PD）。研究目的：探讨PF对MPP+或酸（pH 5.0）诱导的PC12细胞损伤的神经保护作用，并探讨自噬-溶酶体通路（ALP）的活性。阿米洛利（Amiloride, Ami）是一种非选择性酸感离子通道（asic）阻滞剂，由于其在PD啮齿动物模型中具有神经保护作用，因此作为阳性对照药物。材料和方法：用MU法测定细胞活力。乳酸脱氢酶（LDH）法检测细胞损伤程度。采用流式细胞术和Western blot分析细胞凋亡、钙内流和自噬机制。结果：Ami （100 μ M）和PF （50 μ M）均能保护PC12细胞免受MPP+或酸诱导的损伤，MTT试验、乳酸脱氢酶释放和细胞凋亡率均达到较高水平。暴露于MPP+或酸中毒后，胞质游离Ca2+浓度升高，而Ami和PF均减少Ca2+的内流。更重要的是，我们发现Ami和PF的神经保护作用机制与LC3-II蛋白上调密切相关，而LC3-II蛋白与自噬液泡膜特异性相关。此外，MPP+或酸的应用诱导了LAMP2a的过表达，这与伴侣介导的自噬途径的活性直接相关。而Ami和PF抑制了LAMP2a的过表达。结论：我们的数据提供了PF在神经元损伤模型中调节自噬的第一个实验证据，也提供了asic和ALP之间关系的第一个指示。2010爱思唯尔爱尔兰有限公司版权所有。
Ethnopharmacological relevance: Paeoniflorin (PF) is the principal bioactive component of Radix Paeoniae alba, which is widely used in Traditional Chinese Medicine for the treatment of neurodegenerative disorders such as Parkinson's disease (PD).Aim of the study: To evaluate the neuroprotective effects of PF on MPP+- or acid- (pH 5.0) induced injury in cultured PC12 cells and to investigate the activity of autophagy-lysosome pathway (ALP). Amiloride (Ami), a non-selective blocker of acid-sensing ion channels (ASICs), as a positive control drug, since it is neuroprotective in rodent models of PD.Materials and methods: The cell viability was analyzed with MU assay. The cell injury was assessed by lactate dehydrogenase (LDH) assay. Flow cytometry and Western blot analysis were used to study the apoptotic, calcium influx and autophagic mechanisms.Results: Ami (100 mu M) and PF (50 mu M) both protected PC12 cells against MPP+- or acid-induced injury as assessed by MTT assay, lactate dehydrogenase release, and apoptosis rate. The concentrations of cytosolic free Ca2+ were raised after exposure to MPP+ or acidosis, while Ami and PF both reduced the influx of Ca2+. More importantly, we found that the mechanisms of neuroprotective effects of Ami and PF were closely associated with the upregulation of LC3-II protein, which is specifically associated with autophagic vacuole membranes. Furthermore, application of MPP+ or acid induced the overexpression of LAMP2a, which is directly correlated with the activity of the chaperone-mediated autophagy pathway. However, Ami and PF inhibited the overexpression of LAMP2a.Conclusions: Our data provide the first experimental evidence that PF modulates autophagy in models of neuron injury, as well as providing the first indication of a relationship between ASICs and ALP. (C) 2010 Elsevier Ireland Ltd. All rights reserved.