Physalin A induces apoptosis via p53-Noxa-mediated ROS generation, and autophagy plays a protective role against apoptosis through p38-NF-κB survival pathway in A375-S2 cells
Physalin A induces apoptosis via p53-Noxa-mediated ROS generation, and autophagy plays a protective role against apoptosis through p38-NF-κB survival pathway in A375-S2 cells
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DOI：
10.1016/j.jep.2013.04.051
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发表时间：
2013-07-09
影响因子：
5.4
通讯作者：
Ikejima, Takashi
中科院分区：
文献类型：
--
作者：
He, Hao;Zang, Ling-He;Ikejima, Takashi
民族药理学相关性：酸浆素A是一种从天然植物酸浆中分离的生物活性物质。franchetii（Mast.）本研究的目的：在前人研究酸浆素A对人黑色素瘤A375-S2细胞的细胞毒作用的基础上，进一步阐明酸浆素A对人黑色素瘤A375-S2细胞的细胞毒作用的分子机制。材料与方法：MTT法检测A375-S2细胞存活率，相差显微镜、荧光显微镜、siRNA转染、流式细胞术和western blot分析研究酸浆素A诱导A375-S2细胞死亡的机制。我们证明，酸浆素A以时间和剂量依赖性的方式降低存活的A375-S2细胞的比例，并且将A375-S2细胞暴露于酸浆素A导致凋亡和自噬。此外，酸浆素A诱导的细胞凋亡是通过激活p53-Noxa通路和细胞内活性氧（ROS）的产生来触发的。ROS清除剂NAC和GSH的管理导致酸浆素A诱导的ROS产生和细胞凋亡的完全抑制。应用p53抑制剂PFT-α或转染Noxa-siRNA也可以导致相同的结果。自噬通过上调p38-NF-κ B B存活通路发挥抗凋亡作用。另外，用特异性自噬抑制剂3 MA抑制自噬，或用p38抑制剂SB 203580或NF-κ B抑制剂PDTC阻断p38-NF-κ B B通路，均明显促进酸浆素A诱导的细胞凋亡。酸浆素A通过p53-Noxa介导的ROS产生诱导凋亡性细胞死亡，自噬通过上调A375-52细胞p38-NF-κ B存活通路发挥抗凋亡作用。这些结果表明酸浆素A有可能成为未来治疗黑色素瘤的潜在药物。(C)2013爱思唯尔爱尔兰有限公司版权所有。
Ethnopharmacological relevance: Physalin A is a bioactive withanolide isolated from natural plant Physalis alkekengi L var. franchetii (Mast.) Makino, a traditional Chinese herbal medicine named Jindenglong which has long been used for the treatment of cough, sore throat, hepatitis, eczema, dysuria and tumors in China.Aim of the study: Based on the previous study that physalin A induced cytotoxic effect in human melanoma A375-S2 cells, this study was designed to further illustrate the molecular mechanisms underlying.Materials and methods: Cell viability was evaluated in A375-S2 cells by MTT assay, and the mechanisms involved in physalin A-induced A375-S2 cell death were investigated by phase contrast microscopy and fluorescence microscopy, siRNA transfection, flow cytometry and western blot analysis.Results: We demonstrated that physalin A decreased the proportion of viable A375-S2 cells in a time- and dose-dependent manner, and exposure of A375-S2 cells to physalin A led to both apoptosis and autophagy. Moreover, physalin A-induced apoptosis was triggered by activation of p53-Noxa pathway and intracellular reactive oxygen species (ROS) generation. The administration of ROS scavengers NAC and GSH resulted in the complete inhibition of physalin A-induced ROS generation and apoptosis. Application of p53 inhibitor PFT-alpha or transfection with Noxa-siRNA could also lead to the same results. Autophagy, demonstrated by the punctuate distribution of monodansylcadaverine staining, as well as the change of LC3-II/LC3-I proportion and Beclin 1 activation, played a protective role against apoptosis via up-regulation of the p38-NF-kappa B survival pathway in A375-S2 cells. Additionally, inhibition of autophagy by the specific autophagic inhibitor 3MA or blocking the p38-NF-kappa B pathway with p38 inhibitor SB203580 or NF-kappa B inhibitor PDTC obviously promoted physalin A-induced apoptosis.Conclusions: Physalin A induced apoptotic cell death via p53-Noxa-mediated ROS generation, and autophagy played a protective role against apoptosis through up-regulating the p38-NF-kappa B survival pathway in A375-52 cells. These results stated the possibility that physalin A would be a potential agent for the treatment of melanoma in the future. (C) 2013 Elsevier Ireland Ltd. All rights reserved.