Delayed hyperbaric oxygen therapy promotes neurogenesis through reactive oxygen species/hypoxia-inducible factor-1α/β-catenin pathway in middle cerebral artery occlusion rats.
OBJECTIVE
Hyperbaric oxygen (HBO) has been reported to be neuroprotective and to improve neurofunctional outcomes in acute stroke. However, it is not clear whether delayed HBO enhances endogenous neurogenesis and promotes neurofunctional recovery. The aim of this study is to evaluate the effects of delayed HBO therapy on neurogenesis and its potential mechanisms.


METHODS
One hundred eleven male Sprague-Dawley rats that survived for 7 days from 2 hours of middle cerebral artery occlusion and reperfusion were used. Delayed and multiple HBO were administrated beginning at 7 days after middle cerebral artery occlusion and lasting for 42 days with 3 HBO-free intervals (5 days each). Motor sensory deficits were measured by foot-fault test, and learning and memory abilities were evaluated by Morris water maze. Neurogenesis was examined by double immunostaining of bromodeoxyuridine and doublecortin, bromodeoxyuridine and neuronal nuclei at day 42. For mechanism studies, inhibitors for reactive oxygen species (ROS), hypoxia-inducible factor (HIF)-1α, and β-catenin were administrated, and the levels of ROS, HIF-1α, β-catenin, lymphoid enhancer-binding factor-1, T-cell factor-1, neurogenin-1, doublecortin, and synapsin-1 were assessed by ELISA or Western blot at day 14.


RESULTS
Delayed HBO treatment promoted neurogenesis and improved neurofunctional recovery at day 42, and the improvements were reversed by inhibition of ROS and HIF-1α. Delayed HBO significantly increased ROS and HIF-1α, and upregulated the expression of neurogenin-1, doublecortin, and synapsin-1. Inhibition of ROS and HIF-1α removed the effects of delayed HBO.


CONCLUSIONS
Delayed HBO enhanced endogenous neurogenesis and improved neurofunctional recovery in the late-chronic phase of stroke possibly mediated by ROS/HIF-1α/β-catenin pathway. Delayed HBO may serve as an alternative treatment to improve long-term recovery of stroke survivors.
From the Departments of Physiology and Pharmacology (Q.H., X.L., D.C., Y.C., D.D., Junjia Tang, Jiping Tang, J.H.Z.) and Department of Neurosurgery, Loma Linda University School of Medicine, CA (J.H.Z.).
Qin
Q
Hu
From the Departments of Physiology and Pharmacology (Q.H., X.L., D.C., Y.C., D.D., Junjia Tang, Jiping Tang, J.H.Z.) and Department of Neurosurgery, Loma Linda University School of Medicine, CA (J.H.Z.).
Xiping
X
Liang
From the Departments of Physiology and Pharmacology (Q.H., X.L., D.C., Y.C., D.D., Junjia Tang, Jiping Tang, J.H.Z.) and Department of Neurosurgery, Loma Linda University School of Medicine, CA (J.H.Z.).
Di
D
Chen
From the Departments of Physiology and Pharmacology (Q.H., X.L., D.C., Y.C., D.D., Junjia Tang, Jiping Tang, J.H.Z.) and Department of Neurosurgery, Loma Linda University School of Medicine, CA (J.H.Z.).
Yujie
Y
Chen
From the Departments of Physiology and Pharmacology (Q.H., X.L., D.C., Y.C., D.D., Junjia Tang, Jiping Tang, J.H.Z.) and Department of Neurosurgery, Loma Linda University School of Medicine, CA (J.H.Z.).
Desislava
D
Doycheva
From the Departments of Physiology and Pharmacology (Q.H., X.L., D.C., Y.C., D.D., Junjia Tang, Jiping Tang, J.H.Z.) and Department of Neurosurgery, Loma Linda University School of Medicine, CA (J.H.Z.).
Junjia
J
Tang
From the Departments of Physiology and Pharmacology (Q.H., X.L., D.C., Y.C., D.D., Junjia Tang, Jiping Tang, J.H.Z.) and Department of Neurosurgery, Loma Linda University School of Medicine, CA (J.H.Z.).
Jiping
J
Tang
From the Departments of Physiology and Pharmacology (Q.H., X.L., D.C., Y.C., D.D., Junjia Tang, Jiping Tang, J.H.Z.) and Department of Neurosurgery, Loma Linda University School of Medicine, CA (J.H.Z.). johnzhang3910@yahoo.com.
John H
JH
Zhang
D000818:Animals; D000089662:Doublecortin Protein; D006931:Hyperbaric Oxygenation; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D020244:Infarction, Middle Cerebral Artery / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D008297:Male; D055495:Neurogenesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013997:Time Factors; D051176:beta Catenin / Q000378:metabolism*
C000714848:Dcx protein, rat; D000089662:Doublecortin Protein; C497444:Hif1a protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D017382:Reactive Oxygen Species; D051176:beta Catenin
hyperbaric oxygenation; neurogenesis; reactive oxygen species