芪仙通络方对脑梗死大鼠健侧皮质脊髓束重塑及运动功能恢复的影响及机制  

Effect and Mechanism of Qixian Tongluo Formula on Contralateral Corticospinal Tract Remodeling and Motor Functional Recovery in Rats with Cerebral Infarction

作　　者：周胜强 伍大华[1] 李博 陈彦君 黄佳[3] 王琦 曾雯 谭玲娟 邓奕辉[3] 刘芳[1] ZHOU Shengqiang;WU Dahua;LI Bo;CHEN Yanjun;HUANG Jia;WANG Qi;ZENG Wen;TAN Lingjuan;DENG Yihui;LIU Fang(The Affiliated Hospital of Hunan Academy of Chinese Medicine,Changsha 410006 Hunan,China;The First Hospital of Hunan University of Chinese Medicine,Changsha 410007 Hunan,China;Hunan University of Chinese Medicine,Changsha 410208 Hunan,China;Zhaoqing Hospital,the Third Affiliated Hospital,Sun Yat-Sen University,Zhaoqing 526000 Guangdong,China)

机构地区：[1]湖南省中医药研究院附属医院,湖南长沙410006 [2]湖南中医药大学第一附属医院,湖南长沙410007 [3]湖南中医药大学,湖南长沙410208 [4]中山大学附属第三医院肇庆医院,广东肇庆526000

出　　处：《中药新药与临床药理》2024年第9期1319-1328,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：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)。

摘　　要：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 fun

关 键 词：芪仙通络方 脑梗死 皮质脊髓束 神经重塑 运动功能 大鼠 

分 类 号：R285.5[医药卫生—中药学]