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高压氧对脊髓损伤大鼠MMP9/2表达及运动功能的影响

Effect of hyperbaric oxygen on MMP9/2 expression and motor function in rats with spinal cord injury.

作者信息

Hou Ying-Nuo, Ding Wen-Yuan, Shen Yong, Yang Da-Long, Wang Lin-Feng, Zhang Peng

机构信息

Department of Spine Surgery, The Third Hospital of Hebei Medical University 139 Zi Qiang Road, Shijiazhuang 050051, China.

出版信息

Int J Clin Exp Med. 2015 Sep 15;8(9):14926-34. eCollection 2015.

Abstract

To study the effect of hyperbaric oxygen intervention on the microenvironment of nerve regeneration after spinal cord injury modeling and to explore the possible mechanism of nerve regeneration and functional recovery in rats with spinal cord injury. In 98 adult female SD rats, 90 successful models were obtained, which were divided into sham group, spinal cord injury group and hyperbaric oxygen group using randomized block method, 30/group. Spinal cord injury rat model was established in accordance with the modified Allen method. Motor function was assessed at the time points of before modeling, one day, three days, one week, two weeks, three weeks and four weeks after modeling respectively by BBB rating, inclined plane test and improved Tarlov score. At 3 days after modeling, apoptosis of neuronal cells in spinal cord injury region in experimental group was detected by TUNEL method; gene and protein expression of MMP9/2 in spinal cord injury and surrounding tissues was detected by RT-PCR and Western blot assay. At 4 weeks after modeling, histopathological morphological changes in spinal cord injury were observed by HE staining; fluorogold retrograde tracing was used to observe the regeneration and distribution of spinal cord nerve fibers and axon regeneration was observed by TEM. The three motor function scores in hyperbaric oxygen group at each time point after two weeks of treatment were significantly increased compared with spinal cord injury group (P < 0.05). At 3 d after modeling, apoptosis index in hyperbaric oxygen group were significantly lower than those in spinal cord injury group (P < 0.05). At 72 h after modeling, compared with spinal cord injury group, MMP9/2 gene and protein expression in hyperbaric oxygen group was significantly lower (P < 0.05). At four weeks after modeling, fluorogold positive nerve fibers were the most sham group, followed by hyperbaric oxygen group and spinal cord injury group in order; the differences among the groups were statistically significant (P < 0.05). Under TEM, newborn unmyelinated and myelinated nerve fibers could be observed in the middle cross-section in the sham group and hyperbaric oxygen group; unmyelinated and myelinated nerve fibers in hyperbaric oxygen group were more than those in spinal cord injury group. Hyperbaric oxygen therapy played a protective effect on spinal cord injury through reducing apoptosis of neuronal cells and expression of MMP9/2 gene and protein in rats with spinal cord injury.

摘要

研究高压氧干预对脊髓损伤建模后神经再生微环境的影响,探讨脊髓损伤大鼠神经再生及功能恢复的可能机制。选取98只成年雌性SD大鼠,成功造模90只,采用随机区组法分为假手术组、脊髓损伤组和高压氧组,每组30只。按照改良Allen法建立脊髓损伤大鼠模型。分别于建模前、建模后1天、3天、1周、2周、3周和4周,通过BBB评分、斜板试验和改良Tarlov评分评估运动功能。建模后3天,采用TUNEL法检测实验组脊髓损伤区域神经元细胞凋亡情况;采用RT-PCR和Western blot法检测脊髓损伤及周围组织中MMP9/2的基因和蛋白表达。建模后4周,采用HE染色观察脊髓损伤的组织病理学形态变化;采用荧光金逆行示踪法观察脊髓神经纤维的再生和分布情况,采用透射电镜观察轴突再生情况。治疗2周后,高压氧组各时间点的三项运动功能评分均较脊髓损伤组显著升高(P<0.05)。建模后3天,高压氧组的凋亡指数显著低于脊髓损伤组(P<0.05)。建模后72小时,与脊髓损伤组相比,高压氧组MMP9/2基因和蛋白表达显著降低(P<0.05)。建模后4周,荧光金阳性神经纤维数量假手术组最多,其次为高压氧组和脊髓损伤组,组间差异有统计学意义(P<0.05)。透射电镜下,假手术组和高压氧组脊髓损伤中段横切面可见新生无髓鞘和有髓鞘神经纤维;高压氧组无髓鞘和有髓鞘神经纤维数量多于脊髓损伤组。高压氧治疗可通过减少脊髓损伤大鼠神经元细胞凋亡及MMP9/2基因和蛋白表达,对脊髓损伤起到保护作用。

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