补阳还五汤对大鼠脊髓损伤修复与运动功能康复的作用研究

Effects of Buyang Huanwu Decoction on Motor Functional Recovery after Spinal Cord Injury in Rats

目的 探讨中医补阳还五汤对大鼠脊髓损伤后脊髓组织星形胶质细胞胶质纤维酸性蛋白（glial fibrillary acidic protein,GFAP）和巢蛋白（Nestin）表达的影响及其促进脊髓损伤修复的作用机制。方法 大鼠随机分为假手术组（Sham组）、模型组（Model组）、补阳还五汤低剂量组（L-BYHWD组）、补阳还五汤中剂量组（M-BYHWD组）和补阳还五汤高剂量组（H-BYHWD组）。Model组、补阳还五汤低、中、高4组大鼠行脊髓左侧半横断损伤;模型组不行药物干预,补阳还五汤低、中、高3组分别使用不同剂量补阳还五汤连续灌胃。用免疫组织化学法检测不同组大鼠经补阳还五汤治疗后不同时间段损伤区脊髓组织内GFAP和Nestin表达情况,BBB评分评估大鼠后肢运动功能。结果 补阳还五汤干预后大鼠BBB评分明显高于同时期Model组大鼠（P〈0.05）。脊髓损伤后大鼠脊髓组织GFAP表达出现明显上调,补阳还五汤干预后GFAP表达出现下调。M-BYHWD组和H-BYHWD组大鼠脊髓组织GFAP表达明显低于同时期Model组（P〈0.05）;H-BYHWD组大鼠GFAP表达在术后3～7d明显低于同时期M-BYHWD组（P〈0.05）,而术后14～28d二者无显著差异（P〉0.05）。补阳还五汤干预的脊髓组织Nestin表达水平均高于同时期Model组（P〈0.05）;中、高剂量组Nestin表达水平明显高于同时期低剂量组和Model组（P〈0.05）,而中、高剂量组之间Nestin表达水平无显著差异（P〉0.05）。结论 补阳还五汤能促进大鼠脊髓损伤的修复和运动功能的康复,其作用机制可能与抑制GFAP表达减少瘢痕形成、上调Nestin表达促进神经修复有关。

Objective To study the effects and mechanism of Buyang Huangwu Decoction on rats with spinal cord injury by analyzing the changes of glial fibrillary acidic protein （GFAP） and Nestin expression. Methods SD rats were randomly divided into five groups：sham group, model group, low dose Buyaug Huangwu Decoction group （L-BYHWD）, medium dose Buyang Huangwu Decoction group （M-BYHWD） and high dose Buyang Huangwu Decoction group （H-BYHWD）. Rats of Model group, L-BYHWD group, M-BYHWD group, H-BYHWD group underwent spinal cord hemisection injury;model group without drug intervention, Buyang Huanwu Decoction group were used with different doses of Buyang Huanwu Decoction orally. Immunoehemistry was used to evaluate the expression level of GFAP and Nestin. BBB scales was used to evaluate the motor function of hind limbs in rats. Results The BBB scores of the rats after the intervention of BYHWD was higher than the same period of the model group （P〈0.05）. The expression of GFAP in the rats spinal cord tissue was significantly increased after SCI, while down-regulation in the groups with the intervention of BYHWD. The expression of GFAP in the spinal cord tissue of rats of M-BYHWD and H-BYHWD was significantly lower than the same periods in the model group（P〈0.05 ）; The expression of GFAP in the rats of H-BYHWD significantly lower than the same periods of M-BYHWD in 3-7 days after surgery （P〈0.05）, while it does no significant difference in 28 days after surgery （P〉0.05）. expression of Nestin in the spinal cord tissue with the intervention of BYHWD was higher than the same period model group （P〈0.05）;Both the groups of M-BYHED and H-BYHWD, the expression level of Nestin were significantly higher than the model group in the same period（P〈0.05）, while there was no significant difference of the Nestin expression between the M-BYHWD and the H-BYHWD. Conclusion BYHWD can promote the repair of injured spinal cord in rats and its mechanism may be related to reducing scaring by inhibiting the expression of GFAP and repairing nerve by increasing the expression of Nestin.