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Redox-active antioxidant modulation of lipid signaling in vascular endothelial cells: vitamin C induces activation of phospholipase D through phospholipase A2, lipoxygenase, and cyclooxygenase

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Abstract

We have earlier reported that the redox-active antioxidant, vitamin C (ascorbic acid), activates the lipid signaling enzyme, phospholipase D (PLD), at pharmacological doses (mM) in the bovine lung microvascular endothelial cells (BLMVECs). However, the activation of phospholipase A2 (PLA2), another signaling phospholipase, and the modulation of PLD activation by PLA2 in the ECs treated with vitamin C at pharmacological doses have not been reported to date. Therefore, this study aimed at the regulation of PLD activation by PLA2 in the cultured BLMVECs exposed to vitamin C at pharmacological concentrations. The results revealed that vitamin C (3–10 mM) significantly activated PLA2 starting at 30 min; however, the activation of PLD resulted only at 120 min of treatment of cells under identical conditions. Further studies were conducted utilizing specific pharmacological agents to understand the mechanism(s) of activation of PLA2 and PLD in BLMVECs treated with vitamin C (5 mM) for 120 min. Antioxidants, calcium chelators, iron chelators, and PLA2 inhibitors offered attenuation of the vitamin C-induced activation of both PLA2 and PLD in the cells. Vitamin C was also observed to significantly induce the formation and release of the cyclooxygenase (COX)- and lipoxygenase (LOX)-catalyzed arachidonic acid (AA) metabolites and to activate the AA LOX in BLMVECs. The inhibitors of PLA2, COX, and LOX were observed to effectively and significantly attenuate the vitamin C-induced PLD activation in BLMVECs. For the first time, the results of the present study revealed that the vitamin C-induced activation of PLD in vascular ECs was regulated by the upstream activation of PLA2, COX, and LOX through the formation of AA metabolites involving oxidative stress, calcium, and iron.

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Acknowledgments

This work was supported by the grants from the National Institutes of Health (NIH RO1 NS42617, HL 067176-05, NIDDKD RO1 DK056363, and EB 004031) and the funds from the Dorothy M. Davis Heart and Lung Research Institute and the Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine of the Ohio State University College of Medicine.

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  1. Lipid Signaling and Lipidomics Laboratory, Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Department of Medicine, College of Medicine, The Ohio State University, Columbus, OH, USA

    Emily Steinhour, Shariq I. Sherwani, Jessica N. Mazerik, Valorie Ciapala, Elizabeth O’Connor Butler, Jason P. Cruff, Ulysses Magalang, Sampath Parthasarathy, Chandan K. Sen, Clay B. Marsh, Periannan Kuppusamy & Narasimham L. Parinandi

  2. Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, The Ohio State University, 473 W. 12th Avenue, Columbus, OH, 43210, USA

    Narasimham L. Parinandi

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  1. Emily Steinhour
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Steinhour, E., Sherwani, S.I., Mazerik, J.N. et al. Redox-active antioxidant modulation of lipid signaling in vascular endothelial cells: vitamin C induces activation of phospholipase D through phospholipase A2, lipoxygenase, and cyclooxygenase. Mol Cell Biochem 315, 97–112 (2008). https://doi.org/10.1007/s11010-008-9793-6

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