Effect of Antioxidant Enzymes on Hypoxia-Induced HIF-1α Accumulation and Erythropoietin Activity
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The mechanisms underlying the actions of the antioxidantsupon reactive oxygen species (ROS) generation byNADPH oxidase complex have remained uncertain. In thisstudy, we investigated NADPH oxidase activity and the roleof antioxidant enzymes upon the generation of ROS duringhypoxic stress. ROS generation was found to increase in themouse kidney under hypoxic stress in a time-dependentmanner. Moreover, we found in MCT cells that hypoxiainducedhydrogen peroxide production was decreased byNAC pretreatment. We further analyzed HIF-1α, PHD2and VHL expression in the NAC-pretreated MCT cells andassessed the response of antioxidant enzymes at the transcriptionaland translational levels. SOD3 and Prdx2 weresignificantly increased during hypoxia in the mouse kidney. We also confirmed in hypoxic Prdx2-/-and SOD3 transgenicmice that erythropoietin (EPO) is transcriptionally regulatedby HIF-1α. In addition, although EPO protein was found tobe expressed in a HIF-1α independent manner in threemouse lines, its activity differed markedly between normaland Prdx2-/-/SOD3 transgenic mice during hypoxic stress. Inconclusion, our current results indicate that NADPH oxidasemediatedROS generation is associated with hypoxic stress inthe mouse kidney and that SOD3 and Prdx2 cooperate toregulate cellular redox reactions during hypoxia.
Abstract
The mechanisms underlying the actions of the antioxidantsupon reactive oxygen species (ROS) generation byNADPH oxidase complex have remained uncertain. In thisstudy, we investigated NADPH oxidase activity and the roleof antioxidant enzymes upon the generation of ROS duringhypoxic stress. ROS generation was found to increase in themouse kidney under hypoxic stress in a time-dependentmanner. Moreover, we found in MCT cells that hypoxiainducedhydrogen peroxide production was decreased byNAC pretreatment. We further analyzed HIF-1α, PHD2and VHL expression in the NAC-pretreated MCT cells andassessed the response of antioxidant enzymes at the transcriptionaland translational levels. SOD3 and Prdx2 weresignificantly increased during hypoxia in the mouse kidney. We also confirmed in hypoxic Prdx2-/-and SOD3 transgenicmice that erythropoietin (EPO) is transcriptionally regulatedby HIF-1α. In addition, although EPO protein was found tobe expressed in a HIF-1α independent manner in threemouse lines, its activity differed markedly between normaland Prdx2-/-/SOD3 transgenic mice during hypoxic stress. Inconclusion, our current results indicate that NADPH oxidasemediatedROS generation is associated with hypoxic stress inthe mouse kidney and that SOD3 and Prdx2 cooperate toregulate cellular redox reactions during hypoxia.
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