Zan Xing-Chun, Tang Wei, Li Si-Liang, Gong Li, Li Meng-Xing
College of Acupuncture-moxibustion (Rehabilitation Medical College), Anhui University of Chinese Medicine, Hefei 230012, China.
Zhen Ci Yan Jiu. 2019 Aug 25;44(8):547-53. doi: 10.13702/j.1000-0607.180881.
To observe the effect of electroacupuncture (EA) combined with rehabilitation training on regional cerebral blood flow (rCBF) and angiogenesis in rats with acute cerebral ischemia (ACI), so as to explore its mechanisms underlying improvement of ACI.
A total of 135 male SD rats were divided into 5 groups: sham-operation (sham), model, EA, rehabilitation training and EA+rehabilitation training (combined treatment) groups (=27 rats in each group). The ACI model was established by occlusion of the middle cerebral artery with thread embolus. EA (2 Hz/20 Hz, 3-5 V) was applied to "Baihui" (GV20), "Shuigou" (GV26) and bilateral "Neiguan" (PC6) for 20 min, once daily for 14 days. The rehabilitation training including hair-brushing in an enriched environment (10 min), round wooden-stick turning (10 min), grid-board climbing (10 min), and treadmill running (30 min/d) was condacted once daily for 14 days. The rCBF was measured by Doppler ultrasound. The cerebral infarct volume (CIV) was measured after 2, 3, 5-triphenyltetrazolium chloride (TTC) staining. The expression of CD34 in the ischemic penumbra region of brain tissue was detected by immunohistochemistry, and the expressions of angiogenesis-related factors as vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (VEGFR) and basic fibroblast growth factor (bFGF) proteins in the ischemic brain tissue were detected by Western blot.
Following modeling, the rCBF levels at the 5 min, 3, 7 and 14 day were significantly decreased in the model group relevant to the sham group (<0.01). After the intervention, the rCBF levels were significantly increased on day 3, 7 and 14 in the combined treatment group and on day 7 and 14 in both the EA and rehabilitation training groups in comparison with the model group (<0.01). The CIV was obvious in the model group in comparison with the sham group (<0.01), but was markedly smaller in the EA, rehabilitation training and combined treatment groups on day 3,7 and 14 than in the model group (<0.01). The number of CD34 positive cells, and the expression levels of VEGF, VEGFR, and bFGF proteins in ischemic brain tissues were significantly higher on day 3, 7 and 14 in the model group than in the sham group (<0.01, <0.05), and were further up-regulated considerably at the 3 time-points in the 3 treatment groups (<0.01, <0.05). The therapeutic effect of EA+rehabilitation training was significantly superior to that of simple EA and simple rehabilitation training in up-regulating rCBF, CD34 positive cell number, and expression levels of VEGF, VEGFR and bFGF, and in down-regulating the CIV on day 3,7 and 14 (<0.05, <0.01). No significant differences were found between the EA and rehabilitation groups in the above-mentioned 6 indexes (>0.05).
EA combined with rehabilitation training can reduce the infarct volume and increase rCBF in ACI rats, which is probably associated with its effects in promoting the expression of angiogenesis-related factors of ischemic brain tissues. The effect of EA combined with rehabilitation training is markedly better than that of EA and rehabilitation training alone.
观察电针结合康复训练对急性脑缺血（ACI）大鼠脑局部血流量（rCBF）及血管生成的影响，以探讨其改善ACI的作用机制。
将135只雄性SD大鼠分为5组：假手术组（sham）、模型组、电针组、康复训练组和电针+康复训练联合治疗组（每组27只大鼠）。采用线栓法阻塞大脑中动脉建立ACI模型。电针（2Hz/20Hz，3-5V）刺激“百会”（GV20）、“水沟”（GV26）及双侧“内关”（PC6），每次20分钟，每日1次，共14天。康复训练包括在丰富环境中刷毛（10分钟）、转动圆木棍（10分钟）、攀爬网格板（10分钟）及跑步机跑步（30分钟/天），每日1次，共14天。采用多普勒超声测量rCBF。经2,3,5-氯化三苯基四氮唑（TTC）染色后测量脑梗死体积（CIV）。采用免疫组织化学法检测脑组织缺血半暗带区域CD34的表达，采用蛋白质印迹法检测缺血脑组织中血管内皮生长因子（VEGF）、血管内皮生长因子受体2（VEGFR）和碱性成纤维细胞生长因子（bFGF）等血管生成相关因子蛋白的表达。
建模后，模型组5分钟、3天、7天和14天的rCBF水平与假手术组相比显著降低（P<0.01）。干预后，联合治疗组第3天、7天和14天以及电针组和康复训练组第7天和14天的rCBF水平与模型组相比显著升高（P<0.01）。与假手术组相比，模型组的CIV明显增大（P<0.01），但电针组、康复训练组和联合治疗组在第3天、7天和14天的CIV明显小于模型组（P<0.01）。模型组第3天、7天和14天缺血脑组织中CD34阳性细胞数量以及VEGF、VEGFR和bFGF蛋白的表达水平均显著高于假手术组（P<0.01，P<0.05），且在3个治疗组的这3个时间点均进一步显著上调（P<0.01，P<0.05）。在第3天、7天和14天，电针+康复训练联合治疗在上调rCBF、CD34阳性细胞数量以及VEGF、VEGFR和bFGF的表达水平，下调CIV方面的治疗效果显著优于单纯电针和单纯康复训练（P<0.05，P<0.01）。电针组和康复训练组在上述6项指标上无显著差异（P>0.05）。
电针结合康复训练可减小ACI大鼠的梗死体积并增加rCBF，这可能与其促进缺血脑组织血管生成相关因子表达的作用有关。电针结合康复训练的效果明显优于单纯电针和单纯康复训练。
