丁苯酞动员内皮祖细胞治疗脑缺血再灌注大鼠实验研究

3-n-butylphthalide-mobilized endothelial progenitor cells in treatment of rats following cerebral I/R

目的观察丁苯酞治疗对脑缺血再灌注大鼠的影响。方法选取健康雄性SD大鼠60只,随机分为假手术组、模型组、尼莫地平组[12mg/(kg·d)]和低剂量组[丁苯酞60mg/(kg·d)]和高剂量组[丁苯酞120mg/(kg·d)],线栓法建立大脑中动脉缺血再灌注模型。各组灌胃治疗5d,每天行神经功能缺损评分;流式细胞仪检测缺血2h和治疗5d后循环内皮祖细胞(EPC)水平;行CD31及血管性假血友病因子(vWF)免疫组织化学染色,观察大鼠脑缺血区血管新生情况。结果治疗第2天起,尼莫地平组及高、低剂量组神经功能评分均显著低于模型组(P<0.05),第3天时高剂量组神经功能缺损评分低于低剂量组[(0.40±0.52)分vs(1.10±0.32)分,P<0.05]。假手术组大鼠缺血2h循环EPC水平低于其他4组(P<0.05)。治疗第5天,高剂量组EPC增加的水平明显高于模型组、尼莫地平组和低剂量组(P<0.05)。尼莫地平组及低剂量组和高剂量组脑缺血区CD31阳性及vWF阳性细胞数多于模型组。结论丁苯酞可动员循环EPC,促进缺血区血管新生,改善神经功能,呈剂量依赖性。

Objective To study the effect of 3-n-butylphthalide（NBP）on neurological function,cir culating endothelial progenitor cell （EPC） level and neurogenesis in rats following cerebral I/R. Methods Sixty male SD rats were randomly divided into sham operation group,model group,nimodipine 12 mg/（kg · d） group,low NBP dose 60 mg/（kg · d） group and high NBP dose 120 mg/ （kg · d） group （12 in each group）. A rat MCAO I/R model was established by Longa occlusion. After the animals were treated for 5 days,their neurological function deficits were scored,circulat ing EPC expression was detected by cytometry 2 h after ischemia and 5 days of treatment,CD31 and von Willebrand factor （vWF） were stained with immunohistochemieal method for observing the angiogenesis in cerebral ischemic region of rats. Results The neurological function score was significantly lower in nimodipine group, low NBP dose group and high NBP dose group than in model group on day 2 of treatment （P〈0.05） and significantly lower in high NBP dose group than in low NBP dose group on day 3 of treatment （0.40±0.52 w 1.10±0.32,P〈0.05）. The circulating EPC level was significantly lower in sham operation group than in other groups 2 h after ischemia（P〈0.05） and significantly higher in high NBP dose group than in other groups on day 5 of treatment （P〈0.05）. The number of CD31 and vWF positive cells was greater in nimodipine group, low NBP dose group and high NBP dose group than in model group. Conclusion NBP can mobilize circulating EPC, promote angiogenesis in cerebral ischemic area and improve neurological function in a dose-dependent manner.