电针联合磁刺激治疗对脊髓损伤模型大鼠脊髓AQP-4、MCP-1表达的影响
发布时间：2018-08-19 20:40
【摘要】：目的 1.观察大鼠急性脊髓损伤后和给予电针联合磁刺激后对脊髓组织含水量和炎症的变化,探讨电针和磁刺激对脊髓水肿和炎症反应的影响；2.观察脊髓损伤模型大鼠脊髓中AQP-4和MCP-1分布及表达量的变化；3.进一步观察电针结合磁刺激后AQP-4和MCP-1表达水平的变化,探讨电针结合磁刺激通过调控AQP-4和MCP-1表达治疗脊髓损伤的作用机制。 方法 成年雄性Sprague-Dawley (SD)大鼠50只,体重200±25g,适应性喂养一周后,随机分为电针组、正常组、磁刺激组、模型组和联合治疗组。造模方法：利用重物坠落打击方法制作脊髓损伤动物模型,大鼠以10%水合氯醛按4ml/kg腹腔注射麻醉成功后,将其固定于手术操作台上,背部脱毛备皮消毒,根据浮肋连接的第13椎体作为定位的骨性标志,以此为标志向上数3个胸椎来确定T8棘突的位置,以T8棘突为中心,取后正中线做一纵切口,约3cm,暴露T6-T10椎体,咬除T8棘突和两侧椎板,充分暴露硬脊膜,将10g砝码从3cm高处自由落体撞击停留于硬脊膜上自制的长度和直径均为2mm的探针上,大鼠双下肢及躯体回缩扑动,鼠尾出现痉挛性摆动,示造模成功。正常组和模型组不给予治疗。电针组选取“关元”穴进行电针刺激,选用疏密波,频率2/15Hz,强度lmA,每次20min,每天1次,共治疗10天。磁刺激组在大鼠骶骨处皮肤(刺激S1-S4神经根)上放置直径为40mm的圆形线圈,频率0.5Hz,70%的最大输出强度,每次给予30个刺激冲动,分次完成,每两次刺激中间间隔2分钟,每天治疗1次,共治疗10天。联合治疗组采用电针结合磁刺激治疗,治疗方法、频率、时间同电针组和磁刺激组。治疗结束,用4%多聚甲醛溶液进行心脏灌注内固定,取出的脊髓组织放入4%的多聚甲醛溶液中固定48小时后,采用免疫组化法检测各组大鼠脊髓中AQP-4和MCP-1阳性细胞数量。 结果 模型组大鼠脊髓AQP-4、MCP-1阳性表达密度显著增强,即AQP-4、MCP-1蛋白表达显著增加,与正常组比较具有统计学意义(P0.01)；电针组、磁刺激组和联合治疗组大鼠脊髓AQP-4、MCP-1阳性表达密度也明显增强,即AQP-4、MCP-1蛋白表达明显增加,与正常组比较具有统计学意义(P0.01)。电针组、磁刺激组和联合治疗组大鼠脊髓AQP-4、MCP-1阳性表达密度显著减弱,即AQP-4、MCP-1蛋白表达显著减少,与模型组比较具有统计学意义(P0.01)。电针组和磁刺激组大鼠脊髓AQP-4、MCP-1阳性表达密度显著增强,即AQP-4、MCP-1蛋白表达显著增加,与联合治疗组比较具有统计学意义(P0.01)。电针组与磁刺激组比较,无统计学意义(P0.05)。 结论 电针及磁刺激治疗均能显著降低脊髓损伤后AQP-4、MCP-1含量,减轻脊髓炎症水肿程度,在一定程度上阻止脊髓的继发性损伤,电针结合磁刺激治疗效果优于单纯治疗。
[Abstract]:Objective 1. To observe the changes of water content and inflammation in spinal cord tissue after acute spinal cord injury and electroacupuncture combined with magnetic stimulation in rats, and to explore the effects of electroacupuncture and magnetic stimulation on spinal cord edema and inflammatory response. To observe the distribution and expression of AQP-4 and MCP-1 in spinal cord of rats with spinal cord injury. To observe the changes of AQP-4 and MCP-1 expression after electroacupuncture combined with magnetic stimulation, and to explore the mechanism of electroacupuncture combined with magnetic stimulation in the treatment of spinal cord injury by regulating the expression of AQP-4 and MCP-1. Methods Fifty adult male Sprague-Dawley (SD) rats, weighing 200 卤25 g, were randomly divided into electroacupuncture group, normal group, magnetic stimulation group, model group and combined treatment group. Methods: the animal model of spinal cord injury was made by the method of heavy body falling. The rats were anesthetized with 10% chloral hydrate intraperitoneally according to 4ml/kg, then fixed on the operating table, and disinfected with hair removal and skin preparation on the back. The position of the T8 spinous process was determined by using the 13th vertebrae of the floating rib junction as the bony mark for positioning, and the upper three thoracic vertebrae were used as markers to determine the position of the T8 spinous process. The center line of the T8 spinous process was taken as a longitudinal incision, about 3 cm, to expose the T6-T10 vertebrae. By biting the T8 spinous process and the lamina of both sides, the dural membrane was fully exposed, and the 10g weight was struck from the height of 3cm on the probe with length and diameter of 2mm made by itself on the dura mater, and the lower limbs and the body of the rat were retracted and fluttered. The spastic wobble of rat tail showed that the model was successful. Normal group and model group were not treated. In the electroacupuncture group, "Guanyuan" acupoint was selected for electroacupuncture stimulation. The frequency was 2 / 15 Hz and the intensity was lmA20 min, once a day for 10 days. In the magnetic stimulation group, circular coils with diameter of 40mm were placed on the skin of the sacrum of the rats (stimulation of S1-S4 nerve root). The maximum output intensity of 0.5 Hz was 70%. 30 stimulation impulses were given each time, and the interval between the two stimulations was 2 minutes. The patients were treated once a day for 10 days. The combined treatment group was treated with electroacupuncture combined with magnetic stimulation, the treatment method, frequency and time were the same as the electroacupuncture group and the magnetic stimulation group. The spinal cord tissue was fixed in 4% paraformaldehyde solution for 48 hours. The number of AQP-4 and MCP-1 positive cells in spinal cord of each group was detected by immunohistochemical method. Results the positive expression density of MCP-1 in spinal cord of model group was significantly increased, that is, the expression of MCP-1 protein in AQP-4 was significantly increased compared with that in normal group (P0.01), the expression of MCP-1 in electroacupuncture group was significantly higher than that in normal group (P0.01). The positive expression density of MCP-1 in spinal cord of rats in magnetic stimulation group and combined treatment group was also significantly increased, that is, the expression of MCP-1 protein in AQP-4 was significantly increased compared with that in normal group (P0.01). In electroacupuncture group, magnetic stimulation group and combined treatment group, the positive expression density of AQP-4 and MCP-1 in spinal cord was significantly decreased, that is, the expression of AQP-4 and MCP-1 protein was significantly decreased compared with the model group (P0.01). The positive expression density of MCP-1 in spinal cord of rats in electroacupuncture group and magnetic stimulation group was significantly increased, that is, the expression of MCP-1 protein in AQP-4 was significantly increased compared with that in combined treatment group (P0.01). There was no significant difference between electroacupuncture group and magnetic stimulation group (P0.05). Conclusion both electroacupuncture and magnetic stimulation can significantly reduce the content of AQP-4 and MCP-1 after spinal cord injury, alleviate the degree of inflammation and edema of spinal cord, and to some extent prevent the secondary injury of spinal cord. The effect of electroacupuncture combined with magnetic stimulation is better than that of simple treatment.
【学位授予单位】：湖北中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R245
本文编号：2192772
[Abstract]:Objective 1. To observe the changes of water content and inflammation in spinal cord tissue after acute spinal cord injury and electroacupuncture combined with magnetic stimulation in rats, and to explore the effects of electroacupuncture and magnetic stimulation on spinal cord edema and inflammatory response. To observe the distribution and expression of AQP-4 and MCP-1 in spinal cord of rats with spinal cord injury. To observe the changes of AQP-4 and MCP-1 expression after electroacupuncture combined with magnetic stimulation, and to explore the mechanism of electroacupuncture combined with magnetic stimulation in the treatment of spinal cord injury by regulating the expression of AQP-4 and MCP-1. Methods Fifty adult male Sprague-Dawley (SD) rats, weighing 200 卤25 g, were randomly divided into electroacupuncture group, normal group, magnetic stimulation group, model group and combined treatment group. Methods: the animal model of spinal cord injury was made by the method of heavy body falling. The rats were anesthetized with 10% chloral hydrate intraperitoneally according to 4ml/kg, then fixed on the operating table, and disinfected with hair removal and skin preparation on the back. The position of the T8 spinous process was determined by using the 13th vertebrae of the floating rib junction as the bony mark for positioning, and the upper three thoracic vertebrae were used as markers to determine the position of the T8 spinous process. The center line of the T8 spinous process was taken as a longitudinal incision, about 3 cm, to expose the T6-T10 vertebrae. By biting the T8 spinous process and the lamina of both sides, the dural membrane was fully exposed, and the 10g weight was struck from the height of 3cm on the probe with length and diameter of 2mm made by itself on the dura mater, and the lower limbs and the body of the rat were retracted and fluttered. The spastic wobble of rat tail showed that the model was successful. Normal group and model group were not treated. In the electroacupuncture group, "Guanyuan" acupoint was selected for electroacupuncture stimulation. The frequency was 2 / 15 Hz and the intensity was lmA20 min, once a day for 10 days. In the magnetic stimulation group, circular coils with diameter of 40mm were placed on the skin of the sacrum of the rats (stimulation of S1-S4 nerve root). The maximum output intensity of 0.5 Hz was 70%. 30 stimulation impulses were given each time, and the interval between the two stimulations was 2 minutes. The patients were treated once a day for 10 days. The combined treatment group was treated with electroacupuncture combined with magnetic stimulation, the treatment method, frequency and time were the same as the electroacupuncture group and the magnetic stimulation group. The spinal cord tissue was fixed in 4% paraformaldehyde solution for 48 hours. The number of AQP-4 and MCP-1 positive cells in spinal cord of each group was detected by immunohistochemical method. Results the positive expression density of MCP-1 in spinal cord of model group was significantly increased, that is, the expression of MCP-1 protein in AQP-4 was significantly increased compared with that in normal group (P0.01), the expression of MCP-1 in electroacupuncture group was significantly higher than that in normal group (P0.01). The positive expression density of MCP-1 in spinal cord of rats in magnetic stimulation group and combined treatment group was also significantly increased, that is, the expression of MCP-1 protein in AQP-4 was significantly increased compared with that in normal group (P0.01). In electroacupuncture group, magnetic stimulation group and combined treatment group, the positive expression density of AQP-4 and MCP-1 in spinal cord was significantly decreased, that is, the expression of AQP-4 and MCP-1 protein was significantly decreased compared with the model group (P0.01). The positive expression density of MCP-1 in spinal cord of rats in electroacupuncture group and magnetic stimulation group was significantly increased, that is, the expression of MCP-1 protein in AQP-4 was significantly increased compared with that in combined treatment group (P0.01). There was no significant difference between electroacupuncture group and magnetic stimulation group (P0.05). Conclusion both electroacupuncture and magnetic stimulation can significantly reduce the content of AQP-4 and MCP-1 after spinal cord injury, alleviate the degree of inflammation and edema of spinal cord, and to some extent prevent the secondary injury of spinal cord. The effect of electroacupuncture combined with magnetic stimulation is better than that of simple treatment.
【学位授予单位】：湖北中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R245
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