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目的:观察活血通督汤促进大鼠脊髓损伤后肢体运动功能恢复与脊髓损伤后神经营养因子脑源性神经营养因子(BDNF)、神经生长因子(NGF)和神经营养因子3(NT-3)表达调控的机制。方法:36只SD大鼠随机分为假手术组、模型组、活血通督汤组,每组12只,假手术组仅行椎板切除术,模型组和活血通督汤组均采用NYU脊髓打击器建立脊髓损伤模型,术后第1、3、5、7天行BBB肢体运动功能评分;造模后7d取材,采用尼氏染色光镜观察神经细胞形态,免疫荧光检测BDNF阳性细胞、NGF阳性细胞和NT-3阳性细胞,Western Blot检测BDNF蛋白、NGF蛋白和NT-3蛋白表达情况。结果:(1)活血通督汤可改善脊髓损伤后肢体运动功能,给药第3天起,活血通督汤组BBB评分优于模型组(P<0.05,P<0.01);(2)尼氏染色提示活血通督汤组瘀血面积较小,神经细胞水肿较轻,神经细胞形态较好,空泡样改变较少,损伤神经细胞恢复中,部分神经细胞细胞核肿大,核仁消失。与模型组比较,活血通督汤可增加脊髓损伤后脊髓前角部位BDNF、NGF和NT-3阳性细胞数(P<0.05);与模型组比较,活血通督汤可促进BDNF、NGF和NT-3蛋白表达(P<0.05)。结论:活血通督汤促进脊髓损伤后肢体运动功能恢复的机制之一可能与其上调脊髓灰质前角部位神经营养因子分泌,从而促进脊髓损伤后神经细胞存活、再生和轴突再生、再髓鞘化有关。
OBJECTIVE: To observe the effects of Huoxue Tongsu Tang on the recovery of limb motor function and the expression of neurotrophic factor (BDNF), nerve growth factor (NGF) and neurotrophic factor 3 (NT-3) after spinal cord injury in rats, Mechanisms of expression regulation. Methods: Thirty-six SD rats were randomly divided into sham operation group, model group and Huoxuetongdu Decoction group, 12 rats in each group. The sham operation group was treated by laminectomy only. NYU spinal cord The model of spinal cord injury was established by the beater. The BBB motor function score was performed on the 1st, 3rd, 5th and 7th day after operation. The neurological morphology of the BBB was observed 7 days after the model was made. The BDNF positive cells, NGF Positive cells and NT-3 positive cells were detected by Western Blot BDNF protein, NGF protein and NT-3 protein expression. Results: (1) Huoxue Tong Du Tang improved limb motor function after spinal cord injury. From the third day after administration, the BBB score of Huoxuetongtang group was better than that of the model group (P <0.05, P <0.01); (2) Staining showed that the blood stasis area of Huoxuetongdu Decoction group was smaller, the edema of nerve cells was lighter, the shape of nerve cells was better, the shape of vacuoles was less changed, and in the recovery of injured nerve cells, the nuclei of some nerve cells were swollen and the nucleolus disappeared. Compared with the model group, Huoxue Tongsu Tang increased the number of BDNF, NGF and NT-3 positive cells in the anterior horn of spinal cord after spinal cord injury (P <0.05). Compared with the model group, Huoxuetongtu Decoction could promote BDNF, NGF and NT -3 protein expression (P <0.05). CONCLUSION: One of the mechanisms of Huoxuetongtang promoting limb motor function recovery after spinal cord injury may be related to its up-regulation of neurotrophic factor secretion in the anterior horn of spinal cord to promote neuronal survival, regeneration and axon regeneration after spinal cord injury, remyelination related.