补阳还五汤对大鼠局灶性脑缺血后神经干细胞迁移的影响

目的:观察补阳还五汤对大鼠大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)后神经干细胞(neural stem cells,NSCs)迁移的影响.方法:采用MCAO模型,缺血90 min后再灌.将造模成功的大鼠分层并随机分组分为模型和补阳还五汤组,24 h后ig补阳还五汤12g.kg-1,1次/d,连续3周.另16只不阻塞动脉大鼠设为手术组.分别在给药后1,2,3周进行行为学评分；并在第4天和18天ip 5-溴-2-脱氧尿苷(5-bromo-2-deoxyuridine,BrdU)50 mg· kg -1,各连续3d.2,3,5-三苯基四氮唑(TTC)染色测量梗死面积；免疫组化检测BrdU阳性细胞；酶联免疫法(ELISA)检测基质细胞衍生因子1(stromal cell derived factor-1,SDF-1)蛋白表达水平.结果:与假手术组比较,模型组1周和3周神经行为学评分显著升高(P ＜0.05或P＜ 0.01),脑梗死面积、缺血侧室管膜下区(subventricular zone,SVZ)及纹状体区BrdU阳性细胞数均显著增加(P＜ 0.05或P＜ 0.01).与模型组相比,补阳还五汤能显著改善大鼠MCAO神经功能缺失症状,药后1,2,3周神经功能评分为(1.33±0.60),(o.89±0.58),(0.50±0.31)分,明显低于模型组的(1.72±0.46),(1.56±0.51),(1.89±0.50)分(P＜0.01)；降低脑梗死面积,药后1,3周分别为(16.1±5.2)％,(10.3±0.40)％,明显低于模型组的(35.2±6.3)％,(29.8±6.9)％,(P＜0.01),提高SVZ和纹状体区BrdU阳性细胞数,药后1周分别为(71.17±5.19),(83.8±3.83)个,明显高于模型组的(52.17±6.52),(60.20±6.72)个,药后3周分别为(43.33±6.06),(54.60±5.77)个,明显高于模型组的(35.83±4.88),(22.00±3.87)个(P＜0.01)；给药3周时显著提高SDF-1表达,3周时为(36.3±2.71) pg·mg-1,明显高于模型组的(28.65±3.47)pg·mg-1(P＜ 0.05).结论:补阳还五汤能促进MCAO大鼠缺血侧NSCs迁移,其可能机制是通过上调SDF-1蛋白的表达水平实现的.

Effect of Buyang Huanwu Decoction on Migration of Neural Stem Cells after Focal Cerebral Ischemia in Rats

Objective: To observe the effect of Buyang Huanwu decoction (BYHWD) on migration of neural stem cells after middle cerebral artery occlusion (MCAO) in rats. Method: Cerebral ischemia/reperfusion model of MCAO was established in rats using the suture method, successful modelings were divided into ischemia model group and BYHWD group randomly. BYHWD (12 g·kg^-1) was administered orally 24 h after ischemia once a day for 21 days. Neurological deficit was assessed by neuroethological assessment. The area of cerebral was determined by 2,3,5-triphenyltetrazolium chloride(TTC) staining. 5-bromo-2-deoxyuridine(BrdU) positive cells were examined by immunohistochemistry, the expression of stromal cell derived factor-1(SDF-1) measured by ELISA. Result: After MCAO the neurological behavioral score, cerebral ischemia area, the number of BrdU-positive cells were significantly increased (P<0.05 or P<0.01). Compared to model group, BYHWD could significantly reduce neurological behavioral score and cerebral ischemia areas(P<0.05 or P<0.01), enhance the SDF-1 protein level (P<0.05). The number of BrdU-positive cells in subventricular zone(SVZ) and corpus striatum also increased markedly(P<0.05 or P<0.01). Conclusion: BYHWD improves migration of neural stem cells in ischemic penumbra after MCAO, the possible mechanism is related to the increased expression of SDF-1.