电针联合康复训练对局灶性脑缺血模型大鼠血管新生相关因子bFGF、bFGFR表达的影响

Effect of electroacupuncture combined with rehabilitation training on the expression of bFGF and bFGFR in rats with focal cerebral ischemia

目的:观察电针联合康复训练对大脑中动脉缺血(MCAO)模型大鼠血管新生相关因子碱性成纤维细胞生成因子(bFGF)及其受体(bFGFR)表达的影响,探讨其影响脑内血管新生促进神经恢复的机制。方法:随机选取25只SPF级大鼠为假手术组,其余大鼠复制MCAO模型,造模成功后随机分为模型组、抑制剂组(电针联合康复训练加抑制剂)和针康组(电针联合康复训练),每组25只。电针取穴:百会、风府、内关、心俞。针康组每日予以电针及康复治疗,连续7 d;抑制剂组于每日侧脑室注射蛋白激酶B AKT阻断剂后予以电针及康复治疗,连续7 d。采用改良神经功能损害评分(mNSS)评价大鼠神经功能变化,Western blot检测bFGF、bFGFR蛋白的表达水平,用免疫组织化学法检测CD34+阳性细胞表达,RT-PCR法检测bFGF、bFGFR mRNA表达水平。结果:与假手术组比较,模型组大鼠mNSS评分增高(P<0.01),大鼠缺血侧脑组织bFGF、bFGFR蛋白相对表达量、CD34+阳性细胞表达及bFGF、bFGFR mRNA表达均增高(P<0.01,P<0.05)。与模型组比较,针康组与抑制剂组mNSS评分显著降低(P<0.01),大鼠缺血侧脑组织bFGF、bFGFR蛋白相对表达量、CD34+阳性细胞表达及bFGF、bFGFR mRNA表达均明显增加(P<0.01)。与抑制剂组比较,针康组组大鼠7 d时mNSS评分显著降低(P<0.05),大鼠缺血侧脑组织bFGF、bFGFR蛋白相对表达量、CD34+阳性细胞表达及bFGF、bFGFR mRNA表达均显著降低(P<0.05)。结论:电针联合康复训练通过调控脑缺血后缺血侧血管新生因子bFGF、bFGFR的表达,诱导血管新生,从而促进大鼠神经功能恢复。

Objective:To observe the effect of electroacupuncture combined with rehabilitation training on the expression of angiogenesis-related factors bFGF and bFGFR in rats with middle cerebral artery occlusion model(MCAO)and explore the mechanism of its effect on intracerebral neovascularization for neurological recovery.Methods:25 SPF rats were randomly selected as sham-operated group.The rest rats were replicated with the MCAO model and randomly divided into model group,inhibitor group(electroacupuncture combined with rehabilitation training plus inhibitor)and acupuncture rehabilitation group(electroacupuncture combined with rehabilitation training),with 25 rats in each group.Baihui acupoint(DU20),Fengfu acupoint(DU16),Neiguan acupoint(PC6)and Xinshu acupoint(BL15)were taken by electro-acupuncture.In the acupuncture rehabilitation group,electroacupuncture and rehabilitation were given daily for 7 days.In the inhibitor group,electroacupuncture and rehabilitation were given after daily lateral ventricular injection of AKT blocking agent for 7 days.The modified neurological impairment score(mNSS)was used to evaluate the neurological functional changes in rats.bFGF and bFGFR protein expression levels were detected by Western blot,CD34+positive cell expression was detected by immunohistochemistry,and bFGF and bFGFR mRNA expression levels were detected by RT-PCR.Results:Compared with the sham-operated group,the mNSS score was increased in the model group rats(P<0.01),and the relative expression of bFGF and bFGFR protein,CD34+positive cell expression and bFGF and bFGFR mRNA expression were increased in the brain tissue on the ischemic side of the rats(P<0.01).Compared with the model group,the mNSS score was significantly lower in the acupuncture rehabilitation group and the inhibitor group(P<0.01),and the relative expression of bFGF and bFGFR protein,CD34+positive cell expression and bFGF and bFGFR mRNA expression in the brain tissue on the ischemic side of the rats were significantly increased(P<0.01,P<0.05).Compared with the inhibitor group,the mNSS score was significantly lower(P<0.05)at 7th day.The relative expression of bFGF and bFGFR protein,CD34+positive cell expression and bFGF and bFGFR mRNA expression in brain tissue on the ischemic side of the rats were significantly decreased in the acupuncture rehabilitation group(P<0.05).Conclusion:Electroacupuncture combined with rehabilitation training can induce neovascularization by regulating the expression of bFGF and bFGFR,the ischemic side vascularization factors after cerebral ischemia,thus promoting neurological recovery in rats.