Hydrogen-rich saline can inhibit apoptosis of spinal cord motor neurons in rabbits with spinal cord ischemia-reperfusion injury

doi:10.3969/j.issn.2095-4344.2014.18.013

Abstract

Abstract:

BACKGROUND: Spinal cord ischemia-reperfusion injury is a serious secondary injury of the spinal cord. Multifactor could contribute to the mechanism of this injury, and many therapeutic measures emerge, but the therapeutic effect is not ideal.
OBJECTIVE: To investigate the protective effects and mechanism of hydrogen-rich saline on spinal cord ischemia-reperfusion injury in rabbits.
METHODS: ZIVIN method was adopted to prepare the model of spinal cord ischemia-reperfusion injury. The rabbit models were randomly divided into model group, sham operation group, and hydrogen-rich saline group.
RESULTS AND CONCLUSION: Improved Tarlov scores for the evaluation of motor function were significantly increased in hydrogen-rich saline group compared with the model group at 6, 12, 24, 72 hours after reperfusion (P < 0.01). The contents of malondialdehyde were significantly lower (P < 0.05), while catalase activity was significantly higher (P < 0.05) in hydrogen-rich saline group than that in model group at 72 hours after reperfusion. Hematoxylin-eosin staining revealed that, spinal cord anterior-horn motor neurons maintained intact structure in sham operation group; more necrotic spinal cord anterior-horn motor neurons were found in model group, and granular-vacuolar degeneration occurred in the endochylema. In hydrogen-rich saline group, the structure of spinal cord anterior-horn motor neurons was basically intact, only a small amount of spinal cord anterior-horn motor neurons appeared vacuolar degeneration. TUNEL staining showed no apoptotic spinal cord anterior-horn motor neurons in sham operation group. Many inflammatory cells and apoptotic neurons were found in model group. There were few inflammatory cells and apoptotic neurons in hydrogen-rich saline group. Hydrogen-rich saline can prevent the apoptosis of spinal cord anterior-horn motor neurons in rabbits with spinal cord ischemia-reperfusion injury, and the underlying mechanism is associated with antioxidative effect.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

全文链接：

Key words: hydrogen, spinal cord injuries, reperfusion injury, apoptosis, oxidation-reduction

CLC Number:

R318

Sun Yan-qing, Chen Xiong-sheng, Cao Dong, Zhu Wei, Jia Lian-shun. Hydrogen-rich saline can inhibit apoptosis of spinal cord motor neurons in rabbits with spinal cord ischemia-reperfusion injury[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(18): 2861-2866.

Figures/Tables 2

2.1 实验动物数量分析纳入新西兰白兔30只，实验过程无脱失，均进入结果分析。 2.2 脊髓缺血再灌注损伤兔后肢运动功能变化假手术组兔后肢运动功能正常。缺血再灌注6 h后，模型组、氢盐水组兔后肢运动功能即出现不同程度的损害，再灌注12 h后模型组部分兔后肢运动功能评分再次下降，氢盐水组兔后肢运动功能未见继续恶化。再灌注24 h后模型组兔后肢运动评分全部低于1分，氢盐水组有1只兔后肢运动评分低于2分，其他动物运动功能评分均大于2分。再灌注72 h后模型组兔有5只完全瘫痪，5只仅有可察觉的关节活动，氢盐水组仅有1只无法站立，其他动物运动功能评分均大于3分。经统计分析，再灌注后6，12，24，72 h，氢盐水组后肢运动功能评分明显高于模型组(P < 0.01)。各组动物后肢运动功能评分结果见表1。 2.3 脊髓缺血再灌注损伤兔脊髓组织形态苏木精-伊红染色结果显示，再灌注72 h后光镜下观察假手术组兔脊髓组织灰白质界限清楚，神经细胞形态完整。模型组兔脊髓组织灰白质界限欠清楚，大量神经细胞坏死，胞浆内颗粒变性和空泡变性。氢盐水组神经细胞结构基本完整，核仁明显，见少量神经元细胞空泡变性，见图1。原位末端标记染色结果显示，再灌注72 h后光镜下观察假手术组兔脊髓组织见未见神经细胞凋亡；模型组见脊髓组织破坏严重，大量凋亡的神经细胞，核固缩，核浓染，大量炎性细胞浸润；氢盐水组见脊髓组织少量凋亡细胞伴核固缩、浓染，见图2。 2.4 氢盐水对脊髓组织丙二醛浓度和过氧化氢酶活性的影响再灌注后72 h，与模型组相比，氢盐水组丙二醛降低(P < 0.05)，过氧化氢酶活性升高(P < 0.05)。氢盐水组和模型组丙二醛浓度和过氧化氢酶活性与假手术组相比均差异有显著性意义(P < 0.05)，见表2。"

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