维脑康及银杏总内酯对局灶性脑缺血再灌注自噬和神经发生作用研究

Study on effect of Weinaokang and bilobalide on autophagy and neurogenesis induced by focal cerebral ischemia reperfusion

目的:观察维脑康及其活性成分银杏总内酯对局灶性脑缺血再灌注的保护作用及其机制。方法:采用大鼠60min中脑动脉闭塞(middle cerebral artery occlusion,MCAO)/24 h～14 d再灌注模型。蛋白免疫印迹法检测自噬基因Beclin-1表达。透射电镜观察神经元超微结构变化。免疫荧光法检测缺血侧神经发生。结果:维脑康(20,10 mg.kg-1,ig)及其组分银杏总内酯(10,5 mg.kg-1,ig)可促进缺血侧新生成熟神经元(BrdU+-MAP-2+)产生,抑制自噬基因Beclin-1表达,最终减轻缺血再灌注带来的神经元损伤。结论:维脑康可抑制缺血周边区神经元自噬、促进神经发生,说明自噬可能是有效中药维脑康促进缺血脑区自修复功能的干预靶点。

Objective： To observe the protective effect of the Weinaokang （WNK） and its active compound bilobalide on focal cerebral ischemia reperfusion, and their mechanisms. Method： The 60-minute middle cerebral artery occlusion （MCAO） was adopted to establish the 24 h-14 d reperfusion model The expression of Beclin-1 was detected by the Western blotting technique. The transmis- sion electron microscopy was used to observe ultrastructural changes. Neurogenesis was detected by the immunofluorescence staining. Result： WNK （20, 10 mg . kg-1 , ig） or its active compound bilobalide （ 10, 5 mg . kg-1 , ig） could promote the generation of ma- ture neurons （ BrdU ＋ -MAP-2 ＋ ） at the ischemic side, and inhibit expression of autophagy-related gene Beclin-1, so as to reduce the neuron injury induced by focal cerebral ischemia reperfusion. Conclusion： WNK and its active compound bilobalide can inhibit neuron autophagy and improve neurogenesis in ischemic peripheral area, suggesting that neurogenesis may be the intervention target for WNK to promote self-repairing of ischemic area.