電針聯(lián)合磁刺激治療對(duì)脊髓損傷模型大鼠脊髓AQP-4、MCP-1表達(dá)的影響
發(fā)布時(shí)間：2018-08-19 20:40
【摘要】：目的 1.觀察大鼠急性脊髓損傷后和給予電針聯(lián)合磁刺激后對(duì)脊髓組織含水量和炎癥的變化,探討電針和磁刺激對(duì)脊髓水腫和炎癥反應(yīng)的影響；2.觀察脊髓損傷模型大鼠脊髓中AQP-4和MCP-1分布及表達(dá)量的變化；3.進(jìn)一步觀察電針結(jié)合磁刺激后AQP-4和MCP-1表達(dá)水平的變化,探討電針結(jié)合磁刺激通過(guò)調(diào)控AQP-4和MCP-1表達(dá)治療脊髓損傷的作用機(jī)制。 方法 成年雄性Sprague-Dawley (SD)大鼠50只,體重200±25g,適應(yīng)性喂養(yǎng)一周后,隨機(jī)分為電針組、正常組、磁刺激組、模型組和聯(lián)合治療組。造模方法：利用重物墜落打擊方法制作脊髓損傷動(dòng)物模型,大鼠以10%水合氯醛按4ml/kg腹腔注射麻醉成功后,將其固定于手術(shù)操作臺(tái)上,背部脫毛備皮消毒,根據(jù)浮肋連接的第13椎體作為定位的骨性標(biāo)志,以此為標(biāo)志向上數(shù)3個(gè)胸椎來(lái)確定T8棘突的位置,以T8棘突為中心,取后正中線做一縱切口,約3cm,暴露T6-T10椎體,咬除T8棘突和兩側(cè)椎板,充分暴露硬脊膜,將10g砝碼從3cm高處自由落體撞擊停留于硬脊膜上自制的長(zhǎng)度和直徑均為2mm的探針上,大鼠雙下肢及軀體回縮撲動(dòng),鼠尾出現(xiàn)痙攣性擺動(dòng),示造模成功。正常組和模型組不給予治療。電針組選取“關(guān)元”穴進(jìn)行電針刺激,選用疏密波,頻率2/15Hz,強(qiáng)度lmA,每次20min,每天1次,共治療10天。磁刺激組在大鼠骶骨處皮膚(刺激S1-S4神經(jīng)根)上放置直徑為40mm的圓形線圈,頻率0.5Hz,70%的最大輸出強(qiáng)度,每次給予30個(gè)刺激沖動(dòng),分次完成,每?jī)纱未碳ぶ虚g間隔2分鐘,每天治療1次,共治療10天。聯(lián)合治療組采用電針結(jié)合磁刺激治療,治療方法、頻率、時(shí)間同電針組和磁刺激組。治療結(jié)束,用4%多聚甲醛溶液進(jìn)行心臟灌注內(nèi)固定,取出的脊髓組織放入4%的多聚甲醛溶液中固定48小時(shí)后,采用免疫組化法檢測(cè)各組大鼠脊髓中AQP-4和MCP-1陽(yáng)性細(xì)胞數(shù)量。 結(jié)果 模型組大鼠脊髓AQP-4、MCP-1陽(yáng)性表達(dá)密度顯著增強(qiáng),即AQP-4、MCP-1蛋白表達(dá)顯著增加,與正常組比較具有統(tǒng)計(jì)學(xué)意義(P0.01)；電針組、磁刺激組和聯(lián)合治療組大鼠脊髓AQP-4、MCP-1陽(yáng)性表達(dá)密度也明顯增強(qiáng),即AQP-4、MCP-1蛋白表達(dá)明顯增加,與正常組比較具有統(tǒng)計(jì)學(xué)意義(P0.01)。電針組、磁刺激組和聯(lián)合治療組大鼠脊髓AQP-4、MCP-1陽(yáng)性表達(dá)密度顯著減弱,即AQP-4、MCP-1蛋白表達(dá)顯著減少,與模型組比較具有統(tǒng)計(jì)學(xué)意義(P0.01)。電針組和磁刺激組大鼠脊髓AQP-4、MCP-1陽(yáng)性表達(dá)密度顯著增強(qiáng),即AQP-4、MCP-1蛋白表達(dá)顯著增加,與聯(lián)合治療組比較具有統(tǒng)計(jì)學(xué)意義(P0.01)。電針組與磁刺激組比較,無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論 電針及磁刺激治療均能顯著降低脊髓損傷后AQP-4、MCP-1含量,減輕脊髓炎癥水腫程度,在一定程度上阻止脊髓的繼發(fā)性損傷,電針結(jié)合磁刺激治療效果優(yōu)于單純治療。
[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 鹵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.
【學(xué)位授予單位】：湖北中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R245
本文編號(hào)：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 鹵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.
【學(xué)位授予單位】：湖北中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R245
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