甲泼尼龙琥珀酸钠鞘内注射对大鼠急性脊髓损伤的修复作用及相关蛋白的影响
发布时间：2019-01-18 20:21
【摘要】：采用免疫组织化学方法检测脊髓损伤(spinal cord injury,SCI)后各组不同时段生长相关蛋白-43(Growth associated protein-43,GAP-43)和热休克蛋白27(Heat shock protein27,HSP27)表达情况;各组大鼠在存活期间作后肢Basso Beattie Bresnahan(BBB)运动功能评分;用苏木精-伊红(hematoxylin-eosin,HE)染色法观察损伤的脊髓组织各组不同时段病理变化。以确定甲泼尼龙琥珀酸钠(Methylpredniolone Sodium Succinate,MPss)鞘内注射后是否参与了脊髓神经元内GAP-43和HSP27表达的调节,进一步阐明MPss鞘内注射对急性SCI修复中的可能作用机制。健康成年大鼠体重(210±10)g雌性大鼠90只,随机分为治疗组、观察组、对照组,每组30只。治疗组予以脊髓损伤后MPss 3 mg/kg溶解于0.1 mL 0.9%氯化钠中,鞘内注射;观察组予以单纯手术暴露脊髓,不损伤脊髓;对照组予以脊髓损伤后0.9%氯化钠0.1 m L,鞘内注射。于术后第1,3,7,14,28天进行组织学观察分析、行为学观察。组织学结果:显示GAP-43在大鼠脊髓损伤后均表达增强,观察组有少量表达。治疗组、对照组均出现高表达,在脊髓损伤后1,3,7天表达强度逐渐上升,在第7天达到最高,后表达逐渐下降,对照组第14天降至接近对照组,第28天治疗组、对照组表达均降至相同水平,但仍稍高于观察组。治疗组在早期表达数量及表达强度均高于对照组,随着时间延长,降至与观察组相同水平。图像分析结果显示,治疗组脊髓前角的阳性反应在1,3,7,14d点高于对照组,3d差异最明显,差异有显著性(P0.05)。HSP27在对照组呈低水平表达,治疗组及观察组均在1d达到高峰,治疗组在1,3d的阳性细胞数及阳性染色强度均强于对照组,后表达逐渐下降,治疗组和对照组第7天降至接近观察。治疗组脊髓灰质阳性细胞较对照组高,最明显差别在术后1d,差异有统计学意义(P0.01)。HE染色结果:治疗组、对照组均可见脊髓组织内细胞水肿、结构紊乱、淋巴细胞浸润尼氏体融解等病理变化,治疗组病理改变轻于观察组。观察组脊髓结构正常,无组织内细胞水肿、结构紊乱、淋巴细胞浸润尼氏体融解等病理变化。脊髓组织HE染色结果表明,甲泼尼龙琥珀酸钠鞘内注射可减轻脊髓损伤后的病理改变结果行为学观察:BBB评分提示甲泼尼龙琥珀酸钠鞘内注射可改善大鼠脊髓损伤后后肢运动功能,术后第7d,14d,28d,治疗组、对照组间差异有统计学意义(P0.05),治疗组较对照组恢复明显。甲泼尼龙琥珀酸钠鞘内注射可以促进损伤脊髓神经细胞修复,其作用机制可能与甲泼尼龙琥珀酸钠鞘内注射增强损伤脊髓神经细胞GAP-43和HSP27的表达有关。
[Abstract]:The expression of growth-associated protein-43 (Growth associated protein-43,GAP-43) and heat shock protein (27 (Heat shock protein27,HSP27) were detected by immunohistochemical method after spinal cord injury (spinal cord injury,SCI). The Basso Beattie Bresnahan (BBB) motor function score of hind limbs and the pathological changes of injured spinal cord tissues were observed by hematoxylin-eosin (hematoxylin-eosin,HE) staining. To determine whether intrathecal methylprednisolone sodium succinate (Methylpredniolone Sodium Succinate,MPss) involved in the regulation of GAP-43 and HSP27 expression in spinal cord neurons, and to elucidate the possible mechanism of intrathecal MPss injection in the repair of acute SCI. 90 healthy adult rats weighing (210 卤10) g female rats were randomly divided into treatment group, observation group and control group with 30 rats in each group. After spinal cord injury, MPss 3 mg/kg was dissolved in 0. 1 mL 0.9% sodium chloride and injected intrathecally in the treatment group. The control group was given 0.9% sodium chloride 0.1 mL intrathecal injection after spinal cord injury. Histologic observation and behavioral observation were performed on the 1st day, 3rd day and 1428 th day after operation. Histological results: the expression of GAP-43 was increased after spinal cord injury in rats, and a little in observation group. In the treatment group and the control group, the expression level increased gradually on the 7th day after spinal cord injury, reached the highest level on the 7th day, and then decreased gradually, the control group decreased to close to the control group on the 14th day, and the treatment group on the 28th day. The expression of the control group decreased to the same level, but still slightly higher than the observation group. The quantity and intensity of expression in the treatment group were higher than those in the control group, and decreased to the same level as the observation group. The results of image analysis showed that the positive reaction of spinal cord anterior horn in the treatment group was higher than that in the control group at 714 days after treatment, and the difference was most significant on the 3rd day (P0.05). The expression of HSP27 in the control group was lower than that in the control group. The number of positive cells and the intensity of positive staining in the treatment group were stronger than those in the control group on the 1st day, then the expression decreased gradually, and decreased to close observation on the 7th day in the treatment group and the control group. The positive cells of spinal cord in the treatment group were higher than those in the control group, the most obvious difference was on the 1st day after operation. The difference was statistically significant (P0.01). HE staining result: in the treatment group and the control group, the cell edema and structural disorder in the spinal cord tissue were observed. The pathological changes of lymphocytic infiltration and Nissl body fusion were lighter in the treatment group than in the observation group. In the observation group, the structure of spinal cord was normal, no cell edema, structural disorder, lymphocyte infiltration and melting of Nissl body, and other pathological changes. HE staining of spinal cord tissue showed that, Intrathecal injection of methylprednisolone sodium succinate alleviated the pathological changes after spinal cord injury. BBB score indicated that intrathecal injection of methylprednisolone succinate sodium could improve the motor function of hind limbs in rats with spinal cord injury. The difference between the treatment group and the control group was statistically significant (P0.05), the treatment group was more obvious than the control group. Intrathecal injection of methylprednisolone sodium succinate can promote the repair of injured spinal cord neurons, and its mechanism may be related to the enhancement of the expression of GAP-43 and HSP27 in injured spinal cord neurons by intrathecal injection of methylprednisolone sodium succinate.
【学位授予单位】：河北北方学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
本文编号：2411079
[Abstract]:The expression of growth-associated protein-43 (Growth associated protein-43,GAP-43) and heat shock protein (27 (Heat shock protein27,HSP27) were detected by immunohistochemical method after spinal cord injury (spinal cord injury,SCI). The Basso Beattie Bresnahan (BBB) motor function score of hind limbs and the pathological changes of injured spinal cord tissues were observed by hematoxylin-eosin (hematoxylin-eosin,HE) staining. To determine whether intrathecal methylprednisolone sodium succinate (Methylpredniolone Sodium Succinate,MPss) involved in the regulation of GAP-43 and HSP27 expression in spinal cord neurons, and to elucidate the possible mechanism of intrathecal MPss injection in the repair of acute SCI. 90 healthy adult rats weighing (210 卤10) g female rats were randomly divided into treatment group, observation group and control group with 30 rats in each group. After spinal cord injury, MPss 3 mg/kg was dissolved in 0. 1 mL 0.9% sodium chloride and injected intrathecally in the treatment group. The control group was given 0.9% sodium chloride 0.1 mL intrathecal injection after spinal cord injury. Histologic observation and behavioral observation were performed on the 1st day, 3rd day and 1428 th day after operation. Histological results: the expression of GAP-43 was increased after spinal cord injury in rats, and a little in observation group. In the treatment group and the control group, the expression level increased gradually on the 7th day after spinal cord injury, reached the highest level on the 7th day, and then decreased gradually, the control group decreased to close to the control group on the 14th day, and the treatment group on the 28th day. The expression of the control group decreased to the same level, but still slightly higher than the observation group. The quantity and intensity of expression in the treatment group were higher than those in the control group, and decreased to the same level as the observation group. The results of image analysis showed that the positive reaction of spinal cord anterior horn in the treatment group was higher than that in the control group at 714 days after treatment, and the difference was most significant on the 3rd day (P0.05). The expression of HSP27 in the control group was lower than that in the control group. The number of positive cells and the intensity of positive staining in the treatment group were stronger than those in the control group on the 1st day, then the expression decreased gradually, and decreased to close observation on the 7th day in the treatment group and the control group. The positive cells of spinal cord in the treatment group were higher than those in the control group, the most obvious difference was on the 1st day after operation. The difference was statistically significant (P0.01). HE staining result: in the treatment group and the control group, the cell edema and structural disorder in the spinal cord tissue were observed. The pathological changes of lymphocytic infiltration and Nissl body fusion were lighter in the treatment group than in the observation group. In the observation group, the structure of spinal cord was normal, no cell edema, structural disorder, lymphocyte infiltration and melting of Nissl body, and other pathological changes. HE staining of spinal cord tissue showed that, Intrathecal injection of methylprednisolone sodium succinate alleviated the pathological changes after spinal cord injury. BBB score indicated that intrathecal injection of methylprednisolone succinate sodium could improve the motor function of hind limbs in rats with spinal cord injury. The difference between the treatment group and the control group was statistically significant (P0.05), the treatment group was more obvious than the control group. Intrathecal injection of methylprednisolone sodium succinate can promote the repair of injured spinal cord neurons, and its mechanism may be related to the enhancement of the expression of GAP-43 and HSP27 in injured spinal cord neurons by intrathecal injection of methylprednisolone sodium succinate.
【学位授予单位】：河北北方学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
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本文编号：2411079
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