首乌仙海片对缺血性卒中模型大鼠脑组织GFAP、GAP-43表达的影响

Influences of Shouwuxianhai Tablets on expressions of brain GFAP and GAP-43 in rats with ischemic stroke*

目的研究补益肝肾中药首乌仙海片对大脑中动脉阻塞(MCAO)大鼠星形胶质细胞活性及神经轴突生长的作用。方法12周龄雄性W istar大鼠40只,随机分为假手术组8只、模型组16只和中药组16只。模型组和中药组按Tamura法制成MCAO模型,假手术组大鼠行相同的开颅术而不凝闭大脑中动脉,于造模成功后6 d,中药组灌胃给予首乌仙海片9 g/kg,余2组分别灌胃给予等量生理盐水。假手术组大鼠于灌胃1周后,其余2组大鼠分别于灌胃1周、2周末各处死8只,断头取脑,ABC法染色,采用免疫组化法,进行脑组织胶质纤维酸性蛋白(GFAP)、神经生长相关蛋白质(GAP-43)的测定。结果中药组给药1周时,GFAP的表达较同期模型组与假手术组均有显著增强(P<0.01),中药组给药2周时,GFAP的表达较给药1周时及同期模型组与假手术组均显著增强,差别均有高度统计学意义(P<0.01)。中药组治疗2周时,梗死灶周围区GAP-43表达明显增高,与其他2组比较差别有高度统计学意义(P<0.01)。相关性分析表明,中药组灌胃2周时,GFAP与GAP-43在大鼠脑组织中的表达具有相关性,r=0.75,P=0.031 9,GAP-43表达随GFAP表达的上调而增强。结论首乌仙海片可诱导缺血性卒中后神经功能重塑,与该药改善星形胶质细胞活性,促进神经轴突的再生有关。

Objective To study the effects of Shouwuxianhai Tablets, having the actions of replenishing liver and kidney, on the activity of astrocyte and the growth of axon in rats with middle cerebral artery occlusion （MCAO）. Methods All male Wistar rats （ 12 weeks old, n = 40） were randomly divided into the sham-operation group （n = 8）, model group （n- 16） and Shouwuxianhai Tablets group （ST group, n = 16）. The model of MCAO was established according to Tamura method in the model group and ST group, and the same craniotomy without MCAO was operated in the sham-operation group. After 6 days the ST group was given Shouwuxianhai Tablets （9 g/kg） intragastrically and other two groups were given equal quantity of normal saline solution. There were 8 rats killed in the sham-operation group after one week and 8 respectively in the ST group and model group after one and two weeks. The brain tissue were collected from all groups and stained by ABC staining method. The expressions of glial fibrillary acidic protein （GFAP） and neural growth-associated protein-43 （GAP-43） were determined by using immunohistochemical assay. Results After one week GFAP expression was increased significantly in the ST group compared with that in the model group or sham-operation group （ P 〈 0.01 ）. After two weeks GFAP expression was increased significantly in the ST group compared with that in the model group, sham-operation group or one-week ST group （P 〈 0.01 ）. The difference was very significant statistically. There was no statistical significance in the difference of GAP-43 positive signal density values between model group and sham-operation group. The expression of GAP- 43 increased significantly in the ST group after two weeks compared with that in other two groups and the difference was very significant statistically （P 〈0.01 ）. The correlation analysis showed that GFAP expression was correlated to the expression of GAP- 43 in the ST group after two weeks （ r = 0.75, P = 0.031 9 ） , and the expression of GAP- 43 went up along with the up-regulation of GFAP expression. Conclusion Shouwuxianhai Tablets can induce the remodeling of nerve function after ischemic stroke, which is related to that Shouwuxianhai Tablets can improve the activity of astrocyte and regeneration of axon.