目的 观察芪仙通络方对脑梗死大鼠健侧皮质脊髓束（CST）重塑与运动功能恢复的影响，并从神经重塑 相关调控因子角度探讨其潜在分子机制。方法 采用线栓法复制大脑中动脉阻塞（MCAO）大鼠模型。将 50 只 模型复制成功的大鼠随机分为模型组、胞磷胆碱组（0.054 g·kg-1 ）及芪仙通络方低、中、高剂量组（7.83、15.66、 31.32 g·kg-1 ），每组 10 只，另设假手术组（10 只）。术后第 3 天开始灌胃干预，每日 1 次，连续 26 d。于术后 第 3、14、28 天采用 Longa 评分法评价大鼠粗略运动功能，走横木试验（BWT）评分法评价大鼠精细运动功能； 术后第 14 天健侧运动皮层注射神经示踪剂生物素葡聚糖胺（BDA），顺行示踪皮质脊髓束。术后第 28 天，采用 免疫组化法检测健侧运动皮层及颈髓后索生长相关蛋白 43（GAP-43）、BDA 阳性纤维表达；免疫荧光法检测颈 髓灰质内 BDA 阳性纤维与突触前标志蛋白囊泡谷氨酸转运体 1（VGLUT1）共定位面积；Western Blot 法检测健 侧运动皮层中脑源性神经营养因子（BDNF）、胶质细胞源性神经营养因子（GDNF）、神经生长因子（NGF）以及神 经重塑抑制因子勿动蛋白 A（Nogo-A）、少突胶质细胞髓鞘糖蛋白（OMgp）和髓鞘相关糖蛋白（MAG）的表达； Pearson 相关分析法分析 Longa 评分、BWT 评分与 BDA/VGLUT1 共定位面积的相关性。结果 与假手术组比 较，模型组大鼠运动功能缺损明显，各时间点 Longa 评分明显升高（P＜0.01），BWT 评分明显降低（P＜0.05， P＜0.01），健侧运动皮层及颈髓的 GAP-43 蛋白表达水平明显升高（P＜0.05），颈髓后索越边纤维数量明显增 加（P＜0.05），颈髓灰质内的 BDA/VGLUT1 共定位面积明显增加（P＜0.05），健侧运动皮层中 BDNF、GDNF、 NGF 蛋白表达明显上调（P＜0.05），而 Nogo-A、OMgp、MAG 蛋白表达明显下调（P＜0.05）。与模型组比较， 各给药组大鼠 MCAO 术后第 14、28 天的 Longa 评分均显著降低（P＜0.01），BWT 评分均显著升高（P＜0.01）， 健侧运动皮层及颈髓的 GAP-43 蛋白表达水平均显著升高（P＜0.01），颈髓后索越边纤维数量显著增加（P＜ 0.01），颈髓灰质内的 BDA/VGLUT1 共定位面积均显著增加（P＜0.01），健侧运动皮层中 BDNF、GDNF、NGF 蛋白表达均显著上调（P＜0.05，P＜0.01），而 Nogo-A、OMgp、MAG 蛋白表达均明显下调（P＜0.05），其中高 剂量组作用最明显。Longa 评分与 BDA/VGLUT1 共定位面积显著负相关（r = -0.89，P＜0.01），而 BWT 评分与 BDA/VGLUT1 共定位面积显著正相关（r = 0.84，P＜0.01）。结论 芪仙通络方可促进 MCAO 大鼠脑梗死后健侧 CST 重塑，进而改善运动功能，其分子机制可能与调节健侧运动皮层神经重塑相关调控因子的表达有关。

Objective To observe the effect of Qixian Tongluo Formula on contralateral corticospinal tract （CST） remodeling and motor functional recovery in rats with cerebral infarction， and to explore its potential molecular mechanism from the perspective of regulating factors related to never remodeling. Methods The rat middle cerebral artery occlusion （MCAO） model was established by silk thread ligation. Fifty model rats were randomly divided into model group，citicoline group （0.054 g·kg-1 ），Qixian Tongluo Formula low-，medium- and high- dose （7.83， 15.66， 31.32 g·kg-1 ） groups， and sham operation group， with 10 rats in each group. The intervention administration was started on the 3rd day after operation once a day for 26 consecutive days. On the 3rd，14th and 28th day after operation，the gross motor function was evaluated by Longa score，and the fine motor function was evaluated by beam-walking test （BWT） score. The contralateral motor cortex was injected with the nerve tracer biotin dextran amine （BDA） on the 14 th day after operation to anterogradely trace the CST. On the 28th day after operation， the expression of axonal growth associated protein-43 （GAP-43） and BDA positive fibers in the contralateral motor cortex and cervical spinal cord were detected by immunohistochemistry. The co-localization areas of BDA positive fibers and presynaptic marker protein vesicular glutamate transporter 1 （VGLUT1） in the cervical spinal cord gray matter were detected by immunofluorescence. The expressions of brain-derived neurotrophic factor （BDNF）， glial cell-derived neurotrophic factor （GDNF）， nerve growth factor （NGF） and nerve remodelingassociated inhibitory factor [Nogo-A， oligodendrocyte myelin glycoprotein （OMgp） and myelin-associated glycoprotein （MAG）] in the contralateral motor cortex were detected by Western Blot. Pearson correlation analysis was used to analyze the correlation between Longa score or BWT score and BDA/VGLUT1 co-localization area， respectively. Results Compared with the sham operation group， rats in the model group had obvious symptoms of motor function deficits，and the Longa scores were significantly increased （P＜0.01） and the BWT scores were significantly decreased （P＜0.05， P＜0.01） at each time point. The expression of GAP-43 in the contralateral motor cortex and cervical spinal cord was up-regulated （P＜0.05），the number of edge-crossing fibers from the posterior funiculus in cervical cord was increased （P＜0.05），the co-localization area of BDA/VGLUT1 in the gray matter of the cervical spinal cord was increased （P＜0.05），the expressions of BDNF，GDNF and NGF in the contralateral motor cortex were up-regulated （P＜0.05），while the expressions of Nogo-A，OMgp and MAG were down-regulated （P＜0.05）. Compared with the model group，the Longa scores in each administration group on the 14th and 28th day after MCAO operation were significantly decreased （P＜0.01），the BWT scores were significantly increased （P＜0.01）， the expression of GAP-43 in the contralateral motor cortex and cervical spinal cord was significantly up-regulated （P＜0.01）. The number of edge-crossing fibers from the posterior funiculus in cervical cord was significantly increased （P＜0.01）， the co-localization area of BDA/VGLUT1 in the gray matter of the cervical spinal cord was significantly increased （P＜0.01） . The expressions of BDNF， GDNF and NGF in the contralateral motor cortex were significantly up-regulated （P＜0.01，P＜0.05），while the expressions of Nogo-A， OMgp and MAG were significantly down-regulated （P＜0.05），and the most significant effect was observed in the high dose group. The Longa score was negatively correlated with the co-localization area of BDA/VGLUT1 （r = -0.89，P＜0.01），and the BWT score was positively correlated with the co-localization area of BDA/VGLUT1 （r = 0.84，P＜0.01）. Conclusion Qixian Tongluo Formula can improve motor function through promoting contralateral CST remodeling in MCAO rats after cerebral infarction， and the molecular mechanism may be related to the regulation of the expression of nerve remodeling-associated factor in the contralateral motor cortex.

R285.5

湖南省教育厅资助科研项目（18B253）；湖南省自然科学基金项目（2023JJ30367）；长沙市自然科学基金项目（kq2208144）；湖南省卫生 健康委科研计划项目（D202319019427）；湖南省临床医疗技术创新引导项目（2021SK51010）