益气活血方对脑缺血/再灌注后神经细胞凋亡及相关基因表达的影响

Effects of Qi-reinforcing and blood-activating prescription(益气活血方) on apoptosis of neuronal cells and expression of apoptosis-associated genes after cerebral ischemia/reperfusion

目的:探讨益气活血方对脑缺血/再灌注后神经细胞凋亡及凋亡相关基因B cl-2和F as-L蛋白表达的影响。方法:采用Longa报道的线栓法制备大鼠局灶性脑缺血/再灌注模型。将36只SD大鼠按随机数字表法均分为假手术组、模型组和益气活血方组。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)检测各组大鼠脑组织凋亡神经元细胞;用免疫组化法与医学图像分析相结合的方法检测各组大鼠脑组织B cl-2和F as-L的蛋白表达。结果:①与假手术组相比,模型组大鼠缺血侧脑组织TUNEL阳性细胞及B cl-2、F as-L蛋白阳性细胞数目均增多(P均<0.01);②与模型组相比,益气活血方组大鼠缺血侧脑组织TUNEL阳性细胞及F as-L蛋白阳性细胞数目减少,B cl-2蛋白阳性细胞数目增多(P均<0.01)。结论:益气活血方药(芪丹通脉片)可增强B cl-2蛋白的表达,同时下调F as-L蛋白的表达,从而抑制脑缺血/再灌注后神经细胞凋亡,这可能是益气活血方对脑缺血/再灌注损伤起神经保护作用的机制之一。

Objective： To investigate the effects of Qi-reinforcing and blood-activating prescription （益气活血方） on apoptosis of neuronal cells and expression of apoptosis-associated genes Bcl - 2 and Fas L proteins after cerebral ischemia/reperfusion （I/R）. Methods： According to Longa＇s report, the string inserting method was employed to reproduce the rat model of focal cerebral I/R. Thirty-slx SD rats were randomly divided into sham operation group, model group, Qi-reinforcing and blood-actlvating prescription group. Apoptosis of neuronal cells of rats＇ brain tissues in each group was analyzed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling （TUNEL）. The immunohistochemistry combined with medical image pathological system （MIPS） were used to measure the number and mean grey level of Bcl - 2 and Fas - L positive cells in rats＇ cerebral cortex of each group. Results： ①Compared with sham operation group, the expression of positive neuronal cells of TUNEL, positive cells of Bcl - 2 and Fas L protein expressions were increased in rats＇ ischemic brain tissues of model group （all P〈0.01）.②Compared with model group, the positive neuronal cells of TUNEL and the positive cells with expression of Fas L protein in Qi reiniorcing and blood-activating prescription group were decreased, while the positive cells with expression of Bcl -2 protein were increased （all P〈0.01）. Conclusion： Qi-reinforcing and blood-activating prescription （Qidan Tongmai tablet （芪丹通脉片）] can inhibit neuronal cells apoptosis by improving the expression of Bcl - 2 protein and down-regulating the expression of Fas - L protein, which may be one of the mechanisms of neuroprotective effects of the prescription on cerebral I/R injury.