title: Neferine prevents NF-κB translocation and protects muscle cells from oxidative stress and apoptosis induced by hypoxia.
abstract: 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 (ΔψM) 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-κB, COX-2, HIF-1α, caspase-3, caspase-9, Bcl2, and Bax. The results of this study revealed that pretreatment of the cells with Nef significantly decreased the ΔψM and ROS in the cells subjected to hypoxia. Further, Nef inhibited NF-κB there by inhibiting the expression of its downstream regulator COX-2, while it induced the functional HIF-1α 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. © 2016 BioFactors, 42(4):407-417, 2016.
authors.affiliation: DRDO BU Center for Life Sciences, Bharathiar University, Coimbatore, Tamil Nadu, India.
authors.forename: Rathinasamy
authors.initials: R
authors.lastname: Baskaran
authors.affiliation: Department of Biotechnology, Translational Research Laboratory, School of Biotechnology and Genetic Engineering, Bharathiar University, Coimbatore, Tamil Nadu, India.
authors.forename: Paramasivan
authors.initials: P
authors.lastname: Poornima
authors.affiliation: Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan.|Graduate Institute of Chinese Medical Science, China Medical University, Taichung, Taiwan.|Department of Health and Nutrition Biotechnology, Asia University, Taichung, Taiwan.
authors.forename: Chih Yang
authors.initials: CY
authors.lastname: Huang
authors.affiliation: DRDO BU Center for Life Sciences, Bharathiar University, Coimbatore, Tamil Nadu, India.|Department of Biotechnology, Translational Research Laboratory, School of Biotechnology and Genetic Engineering, Bharathiar University, Coimbatore, Tamil Nadu, India.|Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan.|Department of Health and Nutrition Biotechnology, Asia University, Taichung, Taiwan.
authors.forename: Viswanadha Vijaya
authors.initials: VV
authors.lastname: Padma
mesh_terms: D021581:Active Transport, Cell Nucleus; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D017209:Apoptosis; D044182:Benzylisoquinolines / Q000494:pharmacology*; D020013:Calcium Signaling; D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D051546:Cyclooxygenase 2 / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism*
chemical_list: D000893:Anti-Inflammatory Agents; D044182:Benzylisoquinolines; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; C498143:RELA protein, human; D017382:Reactive Oxygen Species; D051996:Transcription Factor RelA; C057222:neferine; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human
keywords: HIF-1α; apoptosis; hypoxia; neferine; reactive oxygen species