Hydrogen peroxide administered into the rat spinal cord at the level elevated by contusion spinal cord injury oxidizes proteins, DNA and membrane phospholipids, and induces cell death: attenuation by a metalloporphyrin.
Hydrogen peroxide administered into the rat spinal cord at the level elevated by contusion spinal cord injury oxidizes proteins, DNA and membrane phospholipids, and induces cell death: attenuation by a metalloporphyrin.
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DOI：
10.1016/j.neuroscience.2014.10.063
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发表时间：
2015-01-29
期刊：
影响因子：
3.3
通讯作者：
Bao, F.
中科院分区：
文献类型：
--
作者：
Liu, D.;Bao, F.
关键词：
我们先前证明脊髓损伤(SCI)后过氧化氢浓度[H_2O_2]显著升高。本研究探讨了1)SCI升高的[H_2O_2]是否足以诱导氧化和细胞死亡，2)细胞凋亡是否是H_2O_2诱导的细胞死亡的一种途径，以及3)催化抗氧化剂四(4-苯甲酸)卟啉能否逆转H_2O_2诱导的氧化和细胞死亡。通过微管将H_2O_2注入未损伤的大鼠脊髓，以模拟脊髓损伤的情况。蛋白质和DNA氧化、膜磷脂过氧化(MLP)、细胞死亡和凋亡用氧化和凋亡标志物的组织化学和免疫组化染色来表征。在过氧化氢或人工脑脊液(ACSF)暴露于赋形剂或MnTBAP处理组的切片中对染色细胞进行定量。与ACSF暴露的动物相比，SCI升高的[H_2O_2]显著增加了细胞内蛋白质和DNA的氧化，分别增加了3倍和8倍的MLP。微透析法测定细胞外过氧化氢升高的丙二醛含量。我们证明，脊髓损伤升高的[H_2O_2]显著增加了细胞外丙二醛的水平。与ACSF对照组相比，H_2O_2还显著增加了细胞损失率和TUNEL阳性神经元和活性caspase-3阳性神经元的数量，分别增加了2.3、2.8和5.6倍。我们的结果直接和明确地证明，脊髓损伤后升高的[H_2O_2]参与了脊髓损伤后MLP、蛋白质和DNA的氧化，从而诱导细胞死亡。因此，我们得出结论：1)H_2O_2在继发性脊髓损伤中起促进氧化和促进细胞死亡的作用，2)细胞凋亡是脊髓损伤后[H_2O_2]升高诱导细胞死亡的一个途径，3)caspase激活是H_2O_2诱导脊髓损伤后细胞凋亡的机制之一，4)MnTBAP治疗显著减少了H_2O_2诱导的氧化、细胞损失和细胞凋亡，使之降至ACSF对照组水平，进一步支持了体内证据支持MnTBAP清除H_2O_2的能力。
We previously demonstrated that hydrogen peroxide concentration [H2O2] significantly increases after spinal cord injury (SCI). The present study explored 1) whether SCI-elevated [H2O2] is sufficient to induce oxidation and cell death, 2) if apoptosis is a pathway of H2O2-induced cell death, and 3) whether H2O2-induced oxidation and cell death could be reversed by treatment with the catalytic antioxidant Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP). H2O2 was perfused through a microcannula into the uninjured rat spinal cord to mimic the conditions induced by SCI. Protein and DNA oxidation, membrane phospholipids peroxidation (MLP), cell death and apoptosis were characterized by histochemical and immunohistochemical staining with antibodies against markers of oxidation and apoptosis. Stained cells were quantified in sections of H2O2-, or artificial cerebrospinal fluid (ACSF)-exposed with vehicle-, or MnTBAP-treated groups. Compared with ACSF-exposed animals, SCI-elevated [H2O2] significantly increased intracellular protein and DNA oxidation by three-fold and MLP by eight-fold in neurons, respectively. H2O2-elevated extracellular malondialdehyde was measured by microdialysis sampling. We demonstrrated that SCI-elevated [H2O2] significantly increased extracellular malondialdehyde above pre-injury levels. H2O2 also significantly increased cell loss and the numbers of TUNEL-positive and active caspase-3-positive neurons by 2.3-, 2.8-, and 5.6-fold compared to ACSF controls, respectively. Our results directly and unequivocally demonstrate that SCI-elevated [H2O2] contributes to post-SCI MLP, protein, and DNA oxidation to induce cell death. Therefore, we conclude that 1) the role of H2O2 in secondary SCI is pro-oxidation and pro-cell death, 2) apoptosis is a pathway for SCI-elevated [H2O2] to induce cell death, 3) caspase activation is a mechanism of H2O2-induced apoptosis after SCI, and 4) MnTBAP treatment significantly decreased H2O2-induced oxidation, cell loss, and apoptosis to the levels of ACSF controls, further supporting MnTBAP’s ability to scavenge H2O2 by in vivo evidence.
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