Intermittent hypothermia is neuroprotective in an in vitro model of ischemic stroke.
Intermittent hypothermia is neuroprotective in an in vitro model of ischemic stroke.
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间歇性低温在缺血性中风的体外模型中具有神经保护作用
DOI：
10.7150/ijbs.8868
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发表时间：
2014
影响因子：
9.2
通讯作者：
Pan SY
中科院分区：
文献类型：
--
作者：
Xu SY;Hu YF;Li WP;Wu YM;Ji Z;Wang SN;Li K;Pan SY
目的：采用体外缺氧缺糖(OGD)模型，探讨间歇性低温(IH)对神经元的保护作用及其可能的分子靶点。方法：分离E18d大鼠胚胎皮质神经元，在复氧过程中给予90min的低氧和低温处理后，分别用细胞计数试剂盒(CCK-8)、酶联免疫吸附试验(ELISA)、BCECFAM、Fluo-3 AM、DCFH-DA和DHE、JC-1染色和末端脱氧核苷酸转移酶介导的缺口末端标记法(TUNEL)检测细胞形态、细胞存活率、微管相关蛋白-2(MAP-2)释放、细胞内pH值和钙、活性氧(ROS)生成、线粒体膜电位(Ψm)和神经元死亡。结果：缺氧90min可引起原代神经元形态异常、细胞存活率下降、MAP-2释放、细胞内酸中毒、钙超载、ROS生成增加、Ψm降低和细胞死亡，而持续低温(CH)和间歇低温(IH)可部分抑制这一作用。有趣的是，6h的CH不足以减少细胞内钙超载和稳定线粒体膜电位(Ψm)，而12h的CH能有效地逆转上述变化。所有IH处理(6×1h、4×1.5h或3×2 h)均能有效地减轻细胞内游离钙超载，抑制ROS生成，稳定线粒体膜电位(Ψm)，减少延迟性细胞死亡。然而，只有超过1.5h的IH间期才能有效地防止细胞活力丧失和细胞内pH下降。结论：在体外缺血性卒中模型中，CH和IH均具有神经保护作用，尽管低温持续时间较短，但IH具有与CH相当的神经保护作用。
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.
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