丁基苯酞对脑出血大鼠血管内皮生长因子、血管生成素-2蛋白表达及血管新生的影响 The Effect of Dl-3-n-butylphthalide on Expressions of VEGF, Ang-2 Proteins and Angiogenesis in Intracerebral Hemorrhage Rats

目的 通过观察大鼠脑出血后脑组织中血管内皮生长因子（vaseular endothelial growth factor， VEGF）、血管生成素-2（angiopoietin-2，Ang-2）蛋白在不同时间点的动态表达及不同剂量丁基苯酞干 预后其与新生血管数量的变化，探讨丁基苯酞对脑出血大鼠可能的神经保护作用及机制。 方法 通过自体血注入法制备SD大鼠脑出血模型并随机分为脑出血模型组、丁基苯酞低剂量组 及丁基苯酞中剂量组，以及对照的假手术组。丁基苯酞低、中剂量组大鼠分别予以丁基苯酞10 mg/kg、 25 mg/kg灌胃给药（每日2次），假手术组及脑出血模型组大鼠则在相同时间用同等体积大豆油代替 灌胃。分别在术后1 d、3 d、7 d、15 d评估大鼠神经功能缺损，采用免疫组化法检测各时间点CD34的表 达并进行新生血管计数及血管场面积测定，检测各时间点VEGF、Ang-2蛋白的表达，测定脑出血大鼠 血肿体积。 结果 与脑出血模型组比较，丁基苯酞中剂量组在术后各时间点，低剂量组术后7 d、15 d VEGF蛋 白表达上调；丁基苯酞低、中剂量组术后各时间点An g-2蛋白表达均上调，血管计数均增多；丁基苯酞 低剂量组术后3 d的神经功能缺损评分较低，丁基苯酞中剂量组术后3 d、7 d、15 d的神经功能缺损评 分较低，上述差异均有统计学意义。丁基苯酞低、中剂量组与脑出血模型组，在术后7 d、15 d血肿体 积差异无统计学意义。 结论 丁基苯酞可以显著减轻脑出血大鼠的神经功能缺损，其作用机制可能与上调VEGF、Ang-2蛋 白的表达，增加脑出血血肿周围新生血管密度有关，同时未增加血肿增大的风险。 Abstract: Objective To investigate the possible neuroprotective effect and mechanism of dl-3-nbutylphthalide in intracerebral hemorrhage rats by observing the dynamic expression of vaseular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) protein and the change of neovascularization in brain tissue of rats with intracerebral hemorrhage at different time points after giving different doses of dl-3-n-butylphthalide. Methods Male healthy SD rats were used to prepare cerebral hemorrhage models by injecting autologous blood, and were randomly divided into no-treatment intracerebral hemorrhage model group, the low-dose dl-3-n-butylphthalide group and medium-dose dl-3-n-butylphthalide group, and the sham-operated rats served as control group. Rats in the low and medium dose groups were given dl-3-n-butylphthalide in the dose of 10 mg/kg and 25 mg/kg (twice a day for both groups) respectively, while rats in the sham operation group and no-treatment intracerebral hemorrhage model group were given the same dose of soybean oil at the same time. The following indexes in all groups were assessed at 1, 3, 7 and 15 day after operation: neurological deficit, the expression of CD34, neovascularization and vascular field area, the expressions of VEGF and Ang-2 protein. The volume of hematoma in intracerebral hemorrhage rats were also measured. Results Compared with that in no-treatment model group, the expression of VEGF was up-regulated in the low-dose group at all time points and in the medium-dose group only at 7, 15 days after operation; the expression of Ang-2 protein was up-regulated and the neovascularization count was more in the low-dose and medium-dose group at all time points, and the neurological deficit score was lower in the low-dose group at 3 days after operation and were all lower in the medium-dose group at all time points, and all the above difference had statistical significance. Moreover, there were no significant difference in hematoma volume at 7 and 15 days after operation between the low-dose and medium-dose dl-3-n-butylphthalide groups and no-treatment model group. Conclusions Dl-3-n-butylphthalide can significantly alleviate neurological deficit in cerebral hemorrhage rats, with no increase in the risk of hematoma enlargement. The mechanism of dl-3- n-butylphthalide may be related to up-regulating the expressions of VEGF and Ang-2 protein and increasing the blood vessel density around hematoma.