文章摘要
杨丽芳,文林月,欧小燕,等.颈椎横突尖电项针法调控后循环缺血大鼠血管和神经新生[J].浙江中医药大学学报,2024,48(4):398-406.
颈椎横突尖电项针法调控后循环缺血大鼠血管和神经新生
Cervical Transverse Process Tip Electroacupuncture Therapy Regulates Angiogenesis and Neurogenesis in Rats with Posterior Circulation Ischemia
DOI:10.16466/j.issn1005-5509.2024.04.003
中文关键词: 后循环缺血  针刺  颈椎横突尖电项针法  血管新生  脑缺血  神经发生
英文关键词: posterior circulation ischemia  acupuncture  cervical transverse process tip electroacupuncture therapy  angiogenesis  cerebral ischemia  neurogenesis
基金项目:安徽省高校自然科学研究项目(2023AH050760);全国中医药创新骨干人才培养项目(皖卫中医药发﹝2019﹞12号);安徽省重大疑难疾病中西医协同攻关项目(皖中医药发展秘﹝2021﹞70号);杨骏全国名老中医专家传承工作室项目(国中医药人教发﹝2016﹞42号)
作者单位
杨丽芳 安徽中医药大学 合肥 230031 
文林月 安徽中医药大学 合肥 230031 
欧小燕 安徽中医药大学 合肥 230031 
张寒 广州中医药大学研究生院 
柳刚 安徽中医药大学第二附属医院 
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中文摘要:
      [目的] 观察颈椎横突尖电项针法对后循环缺血大鼠脑血流量和血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)、碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)、内皮抑素(endostatin,ES)和神经巢蛋白(Nestin)、神经元特异性核蛋白(NeuN)蛋白表达的影响,探讨颈椎横突尖电项针法对后循环缺血大鼠血管新生和神经再生的作用及具体机制。[方法] 32只雄性SD大鼠结扎右侧颈总动脉和右锁骨下动脉,复制成永久性后循环缺血模型,假手术组8只仅暴露血管不结扎。将造模成功的24只大鼠分为模型组、西药组和颈椎横突尖电项针组,每组8只。模型组不予任何针刺治疗,但在颈椎横突尖电项针组进行针刺治疗的相同时间内,对该组大鼠进行抓取、捆绑和固定,确保时间、条件与颈椎横突尖电项针组相同;颈椎横突尖电项针组大鼠双侧风池穴和C2、C4、C6颈椎尖位置予以电针治疗;西药组采用尼莫地平溶液灌胃。治疗频率每天一次,连续治疗6 d为1个疗程,1个疗程结束后休息1 d,共2个疗程。采用超声多普勒血流仪检测各组大鼠平均脑血流量,异硫氰酸荧光素-葡聚糖示踪法检测缺血区大脑皮质新生毛细血管数量,苏木精-伊红(hematoxylin-eosin,HE)染色法观察缺血区大脑皮质组织形态,免疫荧光染色检测右侧大脑纹状体中Nestin、NeuN的阳性表达,免疫印迹分析血管新生相关因子VEGF、bFGF、MMP-9、ES蛋白表达水平。[结果] 与假手术组比较,干预后模型组大鼠脑血流量显著降低(P<0.01),新生毛细血管数量增加(P<0.05),右侧纹状体Nestin、NeuN阳性表达增加(P<0.05,P<0.01),缺血区大脑皮质VEGF、MMP-9、bFGF、ES蛋白表达增加(P<0.01,P<0.05)。与模型组比较,西药组和颈椎横突尖电项针组大鼠脑血流量明显上升(P<0.01),缺血区大脑皮质新生毛细血管数量增加(P<0.05),右侧纹状体Nestin、NeuN阳性表达增加(P<0.05,P<0.01),缺血区大脑皮质VEGF、bFGF蛋白表达增加(P<0.05),MMP-9、ES蛋白表达减少(P<0.01)。与西药组比较,颈椎横突尖电项针组大鼠右侧纹状体Nestin、NeuN阳性表达增加(P<0.05),其余指标差异均无统计学意义(P>0.05)。[结论] 颈椎横突尖电项针法可改善后循环缺血大鼠脑血流量,减轻神经缺损程度,促进大鼠血管和神经发生,其作用可能与调控血管新生相关因子的蛋白表达有关。
英文摘要:
      [Objective] To observe the effects of cervical transverse process tip electroacupuncture therapy on cerebral blood flow and protein expression of vascular endothelial growth factor(VEGF), basic fibroblast growth factor(bFGF), matrix metalloproteinase-9(MMP-9), endostatin(ES) and Nestin, NeuN in rats with posterior circulation ischemia, and to investigate the effects of cervical transverse process tip electroacupuncture therapy on angiogenesis and nerve regeneration in rats with posterior circulation ischemia and the specific mechanism of the study. [Methods] Thirty-two male SD rats were replicated into a permanent posterior circulation ischemia model by ligating the right common carotid artery and the right subclavian artery,8 rats in the sham-operated group were only exposed without ligating the vessels, and 24 rats with successful modeling were divided into model group, western medicine group and cervical transverse process tip electroacupuncture group,8 rats in each group. The model group did not receive any acupuncture treatment, but the rats were grasped, bound and fixed at the same time to ensure that the time and conditions were the same as those in cervical transverse process tip electroacupuncture group; the rats in cervical transverse process tip electroacupuncture group were given electroacupuncture treatment to the Fengchi points on the sides and the cervical vertebral apices of the C2, C4 and C6 vertebrae; and western medicine group was treated by gavage with nimodipine solution. The frequency of treatment was 1 time/day,6 consecutive days for 1 course of treatment, and 1 day of rest at the end of 1 course of treatment, for a total of 2 courses of treatment. The mean cerebral blood flow in rats was measured by ultrasonic Doppler flowmetry; the number of new capillaries in the ischemic cerebral cortex of each group of rats was detected by fluorescein isothiocyanate(FITC)-Dextran tracing method; the morphology of cerebral cortex in the ischemic area was observed by hematoxylin-eosin(HE) staining; the positive expression levels of Nestin and NeuN in the right striatum of rat brain was detected by immunofluorescence staining; and the protein expression levels of VEGF, bFGF, MMP-9 and ES were analyzed by Western blot. [Results] After intervention, compared with sham-operated group, model group rats showed significant decrease in cerebral blood flow(P<0.01), increase in the number of new capillaries(P<0.05), increase in the positive expression of Nestin and NeuN in the right striatum(P<0.05, P<0.01), and increase in the protein expression of VEGF, MMP-9, bFGF and ES in the cerebral cortex of the ischemic area(P<0.01, P<0.05). Compared with model group, cerebral blood flow was significantly increased(P<0.01), the intensity of new capillaries in the cerebral cortex of the ischemic area was increased(P<0.05), the positive expression of Nestin and NeuN in the right striatum of the brain was increased(P<0.05, P<0.01), the expression of VEGF and bFGF protein was increased(P<0.01) and the expression of MMP-9 and ES protein was decreased(P<0.01) in the cerebral cortex of the ischemic area in the other two groups. Compared with western medicine group, the positive expression of Nestin and NeuN was increased in the right striatum of rats in cervical transverse process tip electro-acupuncture group(P<0.05), and the rest of the indexes were not statistically significant(P>0.05). [Conclusion] The cervical transverse process tip electroacupuncture therapy can improve cerebral blood flow and the degree of neurological deficit in rats with posterior circulation ischemia and promote angiogenesis and neurogenesis in rats, and its mechanism of action may be related to the regulation of protein expression of factors related to angiogenesis.
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