电针调控大鼠脊髓损伤后灰质中BDNF、NGF和NT3的表达

Electroacupuncture Modulation of the Expressions of BDNF, NGF and NT3 in the Gray Matter After Spinal Cord Injury in Rats

目的观察电针对脊髓损伤后灰质中BDNF、NGF和NT3表达的影响,探讨电针治疗脊髓损伤的相关作用机制。方法将36只SD大鼠随机分为假手术组、模型组、电针组,每组12只。假手术组仅行椎板切除术,模型组和电针组采用脊髓打击器制备脊髓损伤模型。术后电针组给予电针干预,其余各组给予同等条件抓取。术后每日对大鼠进行BBB评分。术后7d取材。采用尼氏染色法观察神经元形态,采用免疫荧光法检测灰质中BDNF、NGF和NT3的表达情况,采用Westernblot检测BDNF蛋白、NGF蛋白和NT3蛋白表达情况,采用实时定量PCR检测BDNFmRNA、NGFmRNA和NT3mRNA的表达量。结果与假手术组比较,模型组和电针组BBB评分显著下降(P<0.05),模型组和电针组灰质中BDNF、NGF和NT3阳性细胞数均显著升高(P<0.05),模型组和电针组BDNF蛋白、NGF蛋白和NT3蛋白表达均显著增加(P<0.05),模型组和电针组BDNFmRNA、NGFmRNA和NT3mRNA表达均显著增加(P<0.05);与模型组比较,电针组BBB评分于干预后5d起显著提高(P<0.05),电针组神经元形态较模型组明显改善,电针组灰质中BDNF、NGF和NT3阳性细胞数均显著升高(P<0.05),电针组BDNF蛋白、NGF蛋白和NT3蛋白表达均显著增加(P<0.05),电针组BDNFmRNA、NGFmRNA和NT3mRNA表达均显著增加(P<0.05)。结论电针能促进大鼠脊髓损伤后灰质中BDNF、NGF和NT3表达,从而有利于神经元修复和大鼠肢体运动功能恢复。

Objective To investigate the effect of electroacupuncture on the expressions of BDNF, NGF and NT3 in the gray matter after spinal cord injury and explore the related mechanism of action of electroacupuncture in treating spinal cord injury. Method Thirty-six SD rats were randomized to sham operation, model and electroacupuncture groups, 12 rats each. The sham operation group only received laminectomy. In the model and electroacupuncture groups, a rat model of spinal cord injury was made using a spinal cord percussion device. After the operation, the electroacupuncture group was given electroacupuncture intervention and the other groups, a grasp under the same conditions. The BBB score was recorded in the rats every day after the operation. The material was taken at 7 days after the operation. Neuron morphology was observed by Nissl staining. The expressions of BDNF, NGF and NT3 in the gray matter were determined by immunofluorescence. The expressions of BDNF protein, NGF protein and NT3 protein were detected by Western blot. The expressions of BDNF mRNA, NGF mRNA and NT3 mRNA were detected by real-time quantitative PCR. Result Compared with the sham operation group, the BBB score decreased significantly (P<0.05), the number of BDNF, NGF and NT3-positive cells in the gray matter increased significantly (P <0.05), the expressions of BDNF protein, NGF protein and NT3 protein increased significantly (P<0.05) and the expressions of BDNF mRNA, NGF mRNA and NT3 mRNA increased significantly (P<0.05) in the model and electroacupuncture groups. Compared with the model group, the BBB score increased significantly from five days after intervention (P<0.05), neuron morphology improved significantly, the number of BDNF, NGF and NT3-positive cells in the gray matter increased significantly (P<0.05), the expressions of BDNF protein, NGF protein and NT3 protein increased significantly (P<0.05) and the expressions of BDNF mRNA, NGF mRNA and NT3 mRNA increased significantly (P<0.05) in the electroacupuncture group. Conclusion Electroacupuncture can promote the expressions of BDNF, NGF and NT3 in the gray matter after spinal cord injury so as to help neuron repair and the restoration of limb motor function in rats.