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目的:研究黄芪加红花对脑缺血再灌注大鼠脑神经细胞凋亡的影响,并通过对缺血周围区caspase-3的检测,进一步探讨黄芪加红花减少神经细胞凋亡的分子机制。方法:采用大鼠脑缺血再灌注模型,分假手术组、模型组、黄芪加红花组、黄芪组、红花组。术后分12 h,24 h,48 h 3个时间点行神经功能评分;TUNEL法观察凋亡的情况;免疫组化观察缺血周围区caspase-3表达强度。结果:神经功能评分显示:与模型组比较,黄芪组、黄芪加红花组神经功能有显著改善;凋亡阳性细胞数:与模型组比较,各治疗组均显著减少;在治疗组中,黄芪加红花组与红花组比较数目减少显著,与黄芪组比较也减少,差别有统计学意义;caspase-3阳性细胞表达水平:缺血再灌注12 h,与模型组比较,黄芪组、黄芪加红花组阳性细胞数显著减少,红花组差别不明显;与红花组比较黄芪加红花组阳性细胞数减少差别显著,与黄芪组比较则差别不显著。缺血再灌注24 h和48 h,治疗组与模型组比较,阳性细胞数均显著减少,黄芪加红花组与红花组、黄芪组比较也显著减少。结论:黄芪加红花为代表的益气活血法比单纯益气或单纯活血具有更好的减轻缺血再灌注后神经细胞凋亡的作用,其作用机制可能通过抑制caspase-3的表达,阻断caspase级联反应的最终执行而实现。
Objective: To study the effects of Astragalus and safflower on neuronal apoptosis in cerebral ischemia-reperfusion rats and to further explore the molecular mechanism of Astragalus and safflower in reducing neuronal apoptosis by detecting the expression of caspase-3 in the peri-ischemic area . Methods: The rat model of cerebral ischemia-reperfusion was established. The rats were divided into sham operation group, model group, Astragalus plus safflower group, Astragalus group and safflower group. Neurological score was evaluated at 12 h, 24 h and 48 h after operation. The apoptosis was observed by TUNEL method. The expression of caspase-3 in the peri-ischemic area was detected by immunohistochemistry. Results: Compared with the model group, the neurological function score showed that the neurological function of the Astragalus membranaceus group and the Radix Astragali group was significantly improved. The number of apoptotic cells: Compared with the model group, the treatment groups were significantly reduced. In the treatment group, Compared with the astragalus group, astragalus membranaceus and Astragalus mongholicus decreased significantly, the expression of caspase-3 positive cells decreased significantly The number of positive cells in safflower group decreased significantly, the difference was not obvious in safflower group; the difference of the number of positive cells between safflower group and safflower group was significant compared with safflower group; but the difference was not significant compared with safflower group. Compared with the model group, the number of positive cells in the treatment group and the model group decreased significantly at 24 h and 48 h after ischemia / reperfusion, and the number of the positive cells in the treated group decreased significantly compared with that in the safflower and astragalus groups. Conclusion: The Qi-invigorating and activating blood-activating method represented by Astragalus mongolica and safflower has the better function of relieving the apoptosis of nerve cells after ischemia-reperfusion than the simple qi-invigorating or simply activating blood circulation. Its mechanism may be through inhibiting the expression of caspase-3, Disruption of caspase cascade reaction to the final implementation.