Chen Wen-Ci, Lin Chu-Yong, Ji Jing, Tu Wen-Zhan, Jiang Song-He
Department of Rehabilitation Medicine, Wenzhou Integrated Chinese and Western Medicine Hospital of Zhejiang Chinese Medical University, Wenzhou 325027, China.
Department of Physical Medicine and Rehabilitation, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325027, Zhejiang Province.
Zhongguo Zhen Jiu. 2021 Mar 12;41(3):307-12. doi: 10.13703/j.0255-2930.20200317-k0005.
To explore the influence of electroacupuncture (EA) on the expression of AMPA receptor subunit GluR1 in the rats with acute spinal cord injury (SCI) and explore the potential effect mechanism of EA in treatment of acute SCI.
A total of 80 SD rats were randomly divided into five groups, i.e. a sham-operation group, a model group, an AMPA antagonist (DNQX) group, an EA group and a DNQX+EA group, 16 rats in each group. The modified Allen's impacting method was adopted to prepare the rat model of acute SCI at T. In the DNQX group, the intrathecal injection of 10 μL DNQX solution with a concentration of 1 nmol/μL was administered in 0.5 h after modeling success. In the EA group, EA (disperse-dense wave, 2 Hz/100 Hz in frequency, 0.5 mA in output current) was given at "Dazhui" (GV 14) and "Mingmen" (GV 4) in 0.5 h, 12 h and 24 h after modeling success for 30 min and totally 3 times. In the DNQX + EA group, the interventions in the above two groups were managed. The Basso, Beattie and Bresnahan locomotor rating score (BBB) was applied to evaluate the changes of locomotor function in the rats before modeling and in 6 h, 24 h and 48 h after modeling successively. Using the hematoxylin-eosin (HE) staining, the histopathological changes in the spinal anterior horn were observed in the spinal injured area. The immunofluorescence method was adopted to determine the number of GluR1 positive neuron of the spinal anterior horn. The Western blot method was used to determine the protein expression of GluR1 in the injured area.
Compared to the sham-operation group in 6 h, 24 h and 48 h after modeling, the BBB scores were all significantly decreased in the model group (<0.001) at the corresponding points. The BBB score was increased in each of intervention groups, but without statistical difference as compared with the model group (>0.05). In the model group, it was found that the boundary between gray matter and white matter in the spinal anterior horn was blurred, the interstitial space enlarged, the neuron volume obviously shrunken, the cytoplasm decreased, the red stain deepened and some neuron nuclei fixed and shrunk. In the EA group, the morphology of the spinal anterior horn in the injured area was improved obviously, which was similar in the DNQX group and the DNQX + EA group. Compared with the sham-operation group, the GluR1 protein expression in the spinal injury area was increased (<0.001) and the number of GluR1 positive neurons elevated (<0.001) in the spinal anterior horn in the model group. Compared with the model group, in the EA group, the DNQX group and the DNQX + EA group, GluR1 protein expression was decreased (<0.05, <0.01) and the number of GluR1 positive neurons in the spinal anterior horn reduced (<0.001).
The intervention with EA at "Dazhui" and "Mingmen" promotes the repair of the injured nerve in the spinal anterior horn probably through inhibiting GluR1 expression in the spinal injured area in the rats with acute SCI.
探讨电针(EA)对急性脊髓损伤(SCI)大鼠脊髓前角α-氨基-3-羟基-5-甲基-4-异恶唑丙酸(AMPA)受体亚基GluR1表达的影响,探讨EA治疗急性SCI的潜在作用机制。
将80只SD大鼠随机分为假手术组、模型组、AMPA拮抗剂(DNQX)组、EA组和DNQX+EA组,每组16只。采用改良Allen打击法制备T10急性SCI大鼠模型。DNQX组于造模成功后0.5 h鞘内注射浓度为1 nmol/μL的DNQX溶液10 μL。EA组于造模成功后0.5 h、12 h和24 h在“大椎”(GV 14)和“命门”(GV 4)穴给予电针(疏密波,频率2 Hz/100 Hz,输出电流0.5 mA),每次30 min,共3次。DNQX+EA组进行上述两组的干预措施。采用Basso、Beattie和Bresnahan运动评分(BBB)评估大鼠造模前及造模成功后6 h、24 h和48 h运动功能变化。采用苏木精-伊红(HE)染色观察脊髓损伤区脊髓前角组织病理学变化。采用免疫荧光法测定脊髓前角GluR1阳性神经元数量。采用蛋白质免疫印迹法检测损伤区GluR1蛋白表达。
与假手术组相比,模型组造模后6 h、24 h和48 h的BBB评分均显著降低(<0.001)。各干预组BBB评分均升高,但与模型组相比无统计学差异(>0.05)。模型组可见脊髓前角灰质与白质界限模糊,间隙增宽,神经元体积明显缩小,细胞质减少,红染加深,部分神经元核固缩。EA组损伤区脊髓前角形态明显改善,DNQX组和DNQX+EA组相似。与假手术组相比,模型组脊髓损伤区GluR1蛋白表达增加(<0.001),脊髓前角GluR1阳性神经元数量增多(<0.001)。与模型组相比,EA组、DNQX组和DNQX+EA组GluR1蛋白表达降低(<0.05,<0.01),脊髓前角GluR1阳性神经元数量减少(<0.001)。
电针“大椎”和“命门”穴干预可能通过抑制急性SCI大鼠脊髓损伤区GluR1表达促进脊髓前角损伤神经修复。