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[电针“大椎”(EX-B2)和“命门”(GV4)对急性脊髓损伤大鼠脊髓前角NR2B表达的影响]

[Influence of electroacupuncture of "Dazhui" (EX-B2) and "Mingmen" (GV4) on NR2B expression in anterior horns of spinal cord in rats with acute spinal cord injury].

作者信息

Chen Wen-Ci, Tu Wen-Zhan, Hu Jie, He Rong, Huang Xiao-Li, Yang Guan-Hu, Jiang Song-He

机构信息

Department of Physical Medicine and Rehabilitation, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325027, Zhejiang Province, China.

Department of Physical Medicine and Rehabilitation, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325027, Zhejiang Province, China; Integrative and Optimized Medicine Research Center, China-USA Institute for Acupuncture and Rehabilitation, Wenzhou Medical University, Wenzhou 325027.

出版信息

Zhen Ci Yan Jiu. 2019 Feb 25;44(2):95-101. doi: 10.13702/j.1000-0607.170970.

Abstract

OBJECTIVE

To observe the influence of eletroacupuncture (EA) at "Dazhui" (EX-B2) and "Mingmen" (GV4) on expression of NR2B subunit of N-methyl-D-aspartate receptor (NMDA) in the injured anterior horn (AH) area of rats with acute spinal cord injury (SCI), so as to explore its mechanisms underlying improvement of neural repair.

METHODS

A total of 96 male Sprague-Dawley (SD) rats were randomly and equally divided into four groups: sham operation (sham), model, medication (Methyl-prednisone, MP) and EA (=24 in each group). The acute SCI model was established by using a MASCIS spinal cord impactor. EA (2 Hz, 0.5 mA) was applied to EX-B2 and GV4 for 30 min, once at 0.5 h, 12 and 24 h after SCI. Rats of the medication group were treated by tail intravenous injection of MP 30 mg/kg within 15 min (impact therapy) and 5.4 mg•kg•h (maintaining treatment) 45 min thereafter for 23 h. The Basso, Beattie and Bresnahan (BBB) rating scale (0 to 21 points) was used to assess changes of locomotor function 6, 24 and 48 h after SCI. Histopathological changes of the injured spinal cord AH region were observed after sectioning and hematoxylin-eosin (H.E.) staining, and the expression levels of NR2B mRNA and protein of AH were measured by quantitative real-time PCR, Western blot and immunofluorescence, respectively.

RESULTS

After SCI, the BBB scores at 6, 24 and 48 h were significantly decreased in the model group compared with those of the sham group (<0.001), but had no remarkable changes in both medication and EA groups after the intervention in comparison with those of the model group (>0.05). After modeling, the histopathological changes (blurred border of the grey-white matter, cellular karyopyknosis, deepening of the cytoplasmic red stain, and rupture, dissolution and disordered arrangement of myelinated nerve fibers) in the injury area of the spinal cord in the model group were apparent, the number of NR2B positive neurons and the relative expression levels of NR2B mRNA and protein were significantly increased in the model group relevant to the sham group (<0.05). In contrast to the model group, the injured severity of the spinal cord AH region was relatively milder, and the expression levels of NR2B mRNA and protein were considerably down-regulated in both EA and MP groups (<0.05). However, there were no significant differences between the EA and MP groups in the expression levels of NR2B mRNA and protein (>0.05).

CONCLUSION

EA at EX-B2 and GV4 may inhibit the expression of NR2B mRNA and protein in acute SCI rats, which may contribute to its action in promoting nerve repair of the injured ventricolumna area of the thoracic spinal cord.

摘要

目的

观察电针“大椎”(EX - B2)、“命门”(GV4)对急性脊髓损伤(SCI)大鼠损伤前角(AH)区N - 甲基 - D - 天冬氨酸受体(NMDA)NR2B亚基表达的影响,以探讨其促进神经修复的机制。

方法

将96只雄性Sprague - Dawley(SD)大鼠随机等分为四组:假手术组(sham)、模型组、药物组(甲泼尼龙,MP)和电针组(每组n = 24)。采用MASCIS脊髓撞击器建立急性SCI模型。电针(2 Hz,0.5 mA)刺激EX - B2和GV4 30 min,于SCI后0.5 h、12 h及24 h各治疗1次。药物组大鼠于SCI后15 min经尾静脉注射MP 30 mg/kg(冲击治疗),45 min后以5.4 mg•kg•h(维持治疗)持续注射23 h。采用Basso、Beattie和Bresnahan(BBB)评分量表(0~21分)于SCI后6 h、24 h及48 h评估运动功能变化。脊髓损伤后行切片及苏木精 - 伊红(H.E.)染色,观察损伤脊髓AH区组织病理学变化,分别采用实时荧光定量PCR、Western blot及免疫荧光法检测AH区NR2B mRNA及蛋白表达水平。

结果

与假手术组比较,模型组SCI后6 h、24 h及48 h的BBB评分显著降低(P < 0.001);与模型组比较,电针组和药物组干预后BBB评分无明显变化(P > 0.05)。造模后,模型组脊髓损伤区组织病理学变化明显(灰白质边界模糊、细胞核固缩、胞质红染加深、有髓神经纤维断裂、溶解及排列紊乱),与假手术组比较,模型组NR2B阳性神经元数量及NR2B mRNA和蛋白相对表达水平显著升高(P < 0.05)。与模型组比较,电针组和MP组脊髓AH区损伤程度相对较轻,NR2B mRNA和蛋白表达水平均明显下调(P < 0.05)。但电针组与MP组NR2B mRNA和蛋白表达水平比较,差异无统计学意义(P > 0.05)。

结论

电针EX - B2和GV4可抑制急性SCI大鼠NR2B mRNA和蛋白表达,这可能是其促进胸段脊髓损伤区神经修复的作用机制之一。

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