Neferine prevents NF-B translocation and protects muscle cells from oxidative stress and apoptosis induced by hypoxia
Neferine prevents NF-B translocation and protects muscle cells from oxidative stress and apoptosis induced by hypoxia
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DOI：
10.1002/biof.1286
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发表时间：
2016-07-01
期刊：
影响因子：
6
通讯作者：
Padma, Viswanadha Vijaya
中科院分区：
文献类型：
--
作者：
Baskaran, Rathinasamy;Poornima, Paramasivan;Padma, Viswanadha Vijaya
甲基莲心碱（Nef）是一种来源于莲子胚的双苄基异喹啉类生物碱，具有广泛的药理活性。Nef在缺氧应激中的细胞保护作用的可能分子机制尚未被探索。因此，这是一个试图阐明参与Nef介导的细胞保护缺氧诱导的细胞损伤的分子机制。用MTT法测定Nef对缺氧的人横纹肌肉瘤细胞的保护作用。500 nM的Nef提供了更好的细胞保护，并用于所有实验。使用荧光探针测量ROS、细胞内钙积累和线粒体膜（M）电位。此外，我们通过流式细胞术和分析NF-B、考克斯-2、HIF-1、caspase-3、caspase-9、Bcl 2和Bax的表达模式来评估Nef对缺氧诱导的炎症和凋亡反应的影响。本研究的结果显示，Nef预处理细胞显著降低缺氧细胞中的M和ROS。此外，Nef通过抑制其下游调节因子考克斯-2的表达来抑制NF-B，同时诱导功能性HIF-1的表达。结果还表明，Nef显着抑制ROS依赖的线粒体介导的缺氧诱导的细胞凋亡。Nef在缺氧诱导的细胞死亡模型中引起的细胞保护作用涉及抗炎和抗凋亡反应。这些结果表明Nef可作为低氧攻击的预防剂。(c)2016年生物因素，42（4）：407-417，2016年
Neferine (Nef), a bisbenzylisoquinoline alkaloid from lotus seed embryo has a wide range of pharmacological activities. Possible molecular mechanism for the cytoprotective action of Nef during hypoxic stress has not been explored till now. Hence, this is an attempt to elucidate the molecular mechanism involved in the Nef mediated cytoprotection on hypoxia-induced cell injury. Cytoprotective dose of Nef in muscle cells (Human rhabdomyosarcoma cells) exposed to hypoxia was determined by MTT assay. Nef at 500 nM offered better cytoprotection and was used for all the experiments. ROS, intracellular calcium accumulation and mitochondrial membrane (M) potential were measured using fluorescent probes. Further, we evaluated the effect Nef on hypoxia induced inflammatory and apoptotic responses by FACS and analyzing the expression patterns of NF-B, COX-2, HIF-1, caspase-3, caspase-9, Bcl2, and Bax. The results of this study revealed that pretreatment of the cells with Nef significantly decreased the M and ROS in the cells subjected to hypoxia. Further, Nef inhibited NF-B there by inhibiting the expression of its downstream regulator COX-2, while it induced the functional HIF-1 expression. The results also indicate that Nef significantly inhibited the ROS dependent mitochondrial mediated apoptosis induced during hypoxia. The cytoprotection elicited by Nef in a model of hypoxia induced cell death involves both anti-inflammatory and anti-apoptotic response. These results suggest that Nef may be used as prophylactic agent against the hypoxic challenge. (c) 2016 BioFactors, 42(4):407-417, 2016