Hypoxia-inducible factor-1α signaling in aquaporin upregulation after traumatic brain injury

Previous studies have demonstrated that traumatic brain injury (TBI) causes brain edema via aquaporins (AQPs), the water-transporting proteins. In the present study, we determined the role of hypoxia inducible factor-1 alpha (HIF-1 alpha), which is a transcription factor in response to physiological hypoxia, in regulating expression of AQP4 and AQP9. Adult male Sprague-Dawley rats (400-425 g) received a closed head injury using the Marmarou weight drop model with a 450 g weight and survived for 1, 4, 24 and 48 h. Some animals were administered 30 min after injury with 2-methoxyestradiol (2ME2), a naturally occurring metabolite of estradiol which is known to post-transcriptionally down-regulate HIF-1 alpha expression, and sacrificed 4h after injury. Real-time PCR and Western blot were used, respectively, to detect gene and protein expressions of manganese superoxide dismutase (MnSOD, showing hypoxic stress), HIF-1 alpha, AQP4, and AQP9. ANOVA analysis demonstrated a significant (p < 0.05) increase in gene expression of MnSOD, HIF-1 alpha, AQP4, and AQP9, starting at I h after injury through 48 h. Western blot analysis further indicated a significant (p < 0.05) increase in protein expression of these molecules at the same time points. Pharmacological inhibition of HIF-1 alpha by 2ME2 reduced the up-regulated levels of AQP4 and AQP9 after TBI. The present study suggests that hypoxic conditions determined by MnSOD expression after closed head injury contribute to HIF-1 alpha expression. HIF-1 alpha, in turn, up-regulates expression of AQP4 and AQP9. These results characterize the pathophysiological mechanisms, and suggest possible therapeutic targets for TBI patients. (C) 2009 Elsevier Ireland Ltd. All rights reserved.