论文部分内容阅读
目的:观察“脑心同治”电针治疗对局灶性脑缺血大鼠血管新生、皮质血管内皮生长因子(VEGF)和脑源性神经营养因子(BDNF)的影响,探讨电针治疗脑缺血的作用机制.方法:共108只Sprague-Dawley大鼠,随机选取27只入假手术组,其余进行右侧大脑中动脉闭塞模型复制后随机分为模型组、传统穴位组及脑心同治组,每组27只.假手术组仅分离颈动脉.传统穴位组电针水沟、曲池、合谷和足三里,脑心同治组电针风府、百会、心俞和内关,均于术后4 h进行,每日1次,连续干预14 d.假手术组、模型组同时间固定,不予任何治疗.治疗前后进行改良神经功能缺损评分(mNSS),检测局部脑血流量(rCBF),免疫组织化学法检测CD34阳性表达,免疫荧光法检测VEGF、BDNF阳性表达,定量反转录聚合酶链式反应(qRT-PCR)法检测VEGF、BDNF mRNA表达.结果:与假手术组比较,模型组mNSS、rCBF及缺血侧皮质微血管密度(MVD)均降低,VEGF、BDNF蛋白及mRNA表达均升高(P<0.01).与模型组比较,两电针组mNSS降低,第14 d时,脑心同治组mNSS较传统穴位组降低(P<0.05);两电针组rCBF较模型组增加,第14 d时rCBF达最高(P<0.05或P<0.01),脑心同治组rCBF高于传统穴位组(P<0.05);7 d、14 d时,两电针组MVD较模型组升高,同时间点脑心同治组MVD高于传统穴位组(P<0.05或P<0.01);两电针组VEGF、BDNF蛋白及mRNA表达较模型组均升高(P<0.01),脑心同治组7 d时VEGF表达最高(P<0.05),7 d、14 d时BDNF表达较3 d时增加(P<0.05),两电针组VEGF、BDNF mRNA表达均在7 d时最高(P<0.05或P<0.01).结论:电针治疗可上调局灶性脑缺血大鼠皮质内VEGF、BDNF表达水平,增加rCBF,这可能是电针治疗脑缺血的作用机制之一;此治疗效果随有效时间累积,其中脑心同治组在促血管新生方面优于传统穴位组.“,”Objective: To observe the effect of electroacupuncture (EA) of “concurrent treatment of the brain and heart” on angiogenesis and cortical vascular endothelial growth factor (VEGF) and brain-derived neurotrophic factor (BDNF) in rats with focal cerebral ischemia, and to explore the mechanism of EA in cerebral ischemia treatment. Methods: A total of 108 Sprague-Dawley rats, 27 rats were randomly selected as the sham-operation group, and the rest rats received the right middle cerebral artery occlusion operation for model preparation firstly, and then were divided into a model group, a traditional acupoint group, and a concurrent treatment of the brain and heart group, with 27 rats in each group. In the sham-operation group, only the carotid artery was isolated. EA at Shuigou (CV26), Quchi (LI11), Hegu (LI4), and Zusanli (ST36) in the traditional acupoint group, and EA at Fengfu (GV16), Baihui (GV20), Xinshu (BL15), and Neiguan (PC6) in the concurrent treatment of the brain and heart group were performed 4 h after the operation, once a day, for 14 consecutive days. Rats in the sham-operation group and the model group were identically fixed without any treatment. Before and after treatment, the modified neurological severity score (mNSS), regional cerebral blood flow (rCBF), and CD34 positive expression by immunohistochemistry were measured. The positive protein expression levels of VEGF and BDNF were detected by immunofluorescence, and the mRNA expression levels of VEGF and BDNF were detected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Results: Compared with the sham-operation group, the mNSS, rCBF, and ischemic side cortical micro-vessel density (MVD) decreased, and the protein and mRNA expression levels of VEGF and BDNF increased in the model group (P<0.01). Compared with the model group, the mNSS of the two EA groups decreased, and the mNSS of the concurrent treatment of the brain and heart group was lower than that of the traditional acupoint group on the 14th day (P<0.05). Compared with the model group, the rCBF in the two EA groups increased, and the rCBF reached the highest on the 14th day (P<0.05 or P<0.01), and the rCBF in the concurrent treatment of the brain and heart group was higher than that in the traditional acupoint group (P<0.05); the MVD of the two EA groups was higher than that of the model group, and the MVD of the concurrent treatment of the brain and heart group was higher than that of the traditional acupoint group on the 7th and 14th days (P<0.05 or P<0.01). Compared with the model group, the protein and mRNA expression levels of VEGF and BDNF in the two EA groups increased (P<0.01). The VEGF expression level was the highest on the 7th day in the concurrent treatment of the brain and heart group (P<0.05), and the BDNF expression level was higher on the 7th and 14th days than on the 3rd day (P<0.05). The mRNA expression levels of VEGF and BDNF in both EA groups reached the highest on the 7th day (P<0.05 or P<0.01). Conclusion: EA therapy can up-regulate the VEGF and BDNF expression levels and increase the rCBF in the cortex of rats with focal cerebral ischemia, which may be one mechanism of EA in the cerebral ischemia treatment. The therapeutic effect is accumulated with the effective time, and the concurrent treatment of the brain and heart group is superior to the traditional acupoint group in promoting angiogenesis.