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

机译:高压氧对脊髓损伤大鼠MMP9 / 2表达和运动功能的影响

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摘要

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.
机译: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,并探讨脊髓损伤大鼠神经再生和功能恢复的可能机制。在98只成年雌性SD大鼠中,成功建立了90只模型,采用随机区组法将其分为假手术组,脊髓损伤组和高压氧组,每组30只。根据改良的Allen方法建立脊髓损伤大鼠模型。在建模前,建模后一天,三天,一周,两周,三周和四周时分别通过BBB评分,斜面测试和改善的Tarlov评分评估运动功能。建模后第3天,采用TUNEL法检测实验组脊髓损伤区域神经元细胞的凋亡。 RT-PCR和Western blot检测MMP9 / 2基因在脊髓损伤及周围组织中的表达。造模后第4周,HE染色观察脊髓损伤的组织病理学形态学变化。氟金逆行示踪法观察脊髓神经纤维的再生和分布,透射电镜观察轴突再生。与脊髓损伤组相比,高压氧组在治疗两周后每个时间点的三个运动功能评分均显着提高(P <0.05)。造模后3 d,高压氧组的细胞凋亡指数明显低于脊髓损伤组(P <0.05)。建模后72 h,高压氧组与脊髓损伤组相比,MMP9 / 2基因和蛋白表达明显降低(P <0.05)。造模后第4周,氟金阳性神经纤维为假手术组,依次为高压氧组和脊髓损伤组。各组之间的差异具有统计学意义(P <0.05)。在透射电镜下,假手术组和高压氧组的中段可观察到新生的无髓和有髓神经纤维。高压氧组的无髓和有髓神经纤维多于脊髓损伤组。高压氧疗法通过减少脊髓损伤大鼠神经元细胞的凋亡以及MMP9 / 2基因和蛋白的表达,对脊髓损伤具有保护作用。

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