通心络治疗缺血再灌注损伤机理的脑片研究

目的 通过器官型脑片研究通心络含药血清对缺血再灌注损伤的治疗作用及机制。方法 制备P3 SD的全脑器官型脑片，随机分为缺血再灌注脑片模型组和正常对照组，各组根据干预方式的不同再分为大剂量含药血清组、小剂量含药血清组、对照血清组、空白组，于干预后不同时间通过倒置显微镜及免疫荧光染色观察通心络对脑片上神经干细胞增殖、迁移、分化的影响。结果 经含药血清干预后室下区、海马齿状回单位区域新生细胞（nestin+）数量增多，正常对照组新生细胞密度高于缺血再灌注脑片模型组（P<0.05）；两组1d、3d、7d时新生细胞数量均按照大剂量含药血清组、小剂量含药血清组、对照血清组、空白组顺序递减（P<0.05）；因新生细胞向外迁移，随着培养时间延长上述区域新生细胞密度逐渐减少；1d时各组均有大量nestin+新生细胞，7d时只有大、小剂量含药血清组有少量nestin+新生细胞。结论 通心络可能通过促进神经干细胞的增殖迁移和分化来治疗缺血再灌注损伤，其效应随剂量增加而增强。

Study on the mechanism of Tongxinluo curing ischemia/reperfusion injury through brain slices

Objective To explore the effect and mechanism of Tongxinluo on curing ischemia/reperfusion injury through organotypic brain slices. Methods Brain slices (300um) from postnatal 3rd day SD rats were prepared and randomly divided into 2 groups: ischemia/reperfusion group and normal group. After enduring ischemia/reperfusion injury, they were divided into 4 groups (high-dose Tongxinluo group, low-dose Tongxinluo group, control group and normal saline group), and administered different reagents. Then they were observed through invert microscope or detected by immunofluorescence staining. Results Neonatal cells labeled with nestin were found in the subventrical zone and hippocampal dentate gyrus when brain slices were administered different dose of Tongxinluo, while ischemia/reperfusion group had less cells (P<0.05). After cultured for 1 day, 3 days and 7 days, the number of neonatal cells decreased in the order of high-dose Tongxinluo group, low-dose Tongxinluo group, control group and normal saline group (p<0.05). The density of neonatal cells in the subventrical zone and hippocampal dentate gyrus decreased due to cell migration as time went on. Each group had lots of neonatal cells labeled with nestin at 1st day, but only two Tongxinluo groups had some neonatal cells labeled with nestin at 7th day. Conclusion Tongxinluo might cure ischemia/reperfusion injury by inducing the proliferation, migration and differentiation of neural stem cells, and the more Tongxinluo was administered the more effects would be achieved.