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International Journal Of Bilogical Sciences

.: Home > International Journal Of Bilogical Sciences > 2014 > Volume 10 Number 8 > Sui-yi Xu1,2, Ya-fang Hu1, Wei-pin Li2, Yong-ming Wu1, Zhong Ji1, Sheng-nan Wang1, Ke Li3, Su-yue Pan1

Intermittent Hypothermia Is Neuroprotective in an in vitro Model of Ischemic Stroke

Sui-yi Xu1,2, Ya-fang Hu1, Wei-pin Li2, Yong-ming Wu1, Zhong Ji1, Sheng-nan Wang1, Ke Li3, Su-yue Pan1
1. Department of Neurology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; 2. Department of Neurosurgery, Shenzhen Key Laboratory of Neurosurgery, Shenzhen Second People’s Hospital, Shenzhen University 1st Affiliated Hospital, Shenzhen 518035, China; 3. Research Center of Clinical Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.  Corresponding author: Dr. Su-yue Pan, Department of Neurology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. Email: pansuyue82@126.com
Abstract :

Objective: To investigate whether the intermittent hypothermia (IH) protects neurons against ischemic insult and the potential molecular targets using an in vitro ischemic model of oxygen glucose deprivation (OGD). Methods: Fetal rat cortical neurons isolated from Day E18 rat embryos were subjected to 90-min OGD and hypothermia treatments during reoxygenation before examining the changes in microscopic morphology, cell viability, microtubule- associated protein 2 (MAP-2) release, intracellular pH value and calcium, reactive oxygen species (ROS) generation, mitochondrial membrane potential (△Ψm) and neuronal death using cell counting kit (CCK-8), enzyme-linked immunosorbent assay (ELISA), BCECF AM, Fluo-3 AM, DCFH-DA and dihydroethidium (DHE), JC-1 staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), respectively. Results: 90-min OGD induced morphologic abnormalities, cell viability decline, MAP-2 release, intracellular acidosis, calcium overload, increased ROS generation, △Ψm decrease and cell death in primary neurons, which was partially inhibited by continuous hypothermia (CH) and intermittent hypothermia (IH). Interestingly, 6-h CH was insufficient to reduce intracellular calcium overload and stabilize mitochondrial membrane potential (△Ψm), while 12-h CH was effective in reversing the above changes. All IH treatments (6×1 h, 4×1.5 h or 3×2 h) effectively attenuated intracellular free calcium overload, inhibited ROS production, stabilized mitochondrial membrane potential (△Ψm) and reduced delayed cell death in OGD-treated cells. However, only IH intervals longer than 1.5 h appeared to be effective in preventing cell viability loss and intracellular pH decline. Conclusion: Both CH and IH were neuroprotective in an in vitro model of ischemic stroke, and in spite of shorter hypothermia duration, IH could provide a comparable neuroprotection to CH.

Keywords :
Ischemic stroke; Hypothermia; Neuroprotection; Primary neuronal culture.

Date Deposited : 12 Feb 2016 10:54

Last Modified : 12 Feb 2016 10:54

Official URL: http://www.ijbs.com/v10i8

Volume 10, Number 8, - 2014 , ISSN 1449-2288

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