目的:探究补阳还五汤对脑出血大鼠脑组织磷脂酰肌醇3-激酶/蛋白激酶B(phosphoinositide 3-kinase/protein kinase B,PI3K/AKT)信号通路、水通道蛋白4(aquaporin-4,AQP4)的影响,阐明作用机制,为脑出血所致脑水肿治疗提供明确的方向与靶点。方法:SD大鼠随机分为6组:假手术组、模型组、补阳还五汤低、中、高剂量组、银杏叶片组,采用Rosenberg法复制大鼠脑出血模型,免疫组化法检测PI3K、AKT蛋白表达,免疫荧光法检测水通道蛋白AQP4"极性"表达,分光光度法检测脑组织伊文思蓝(Evans Blue,EB)含量,干湿重法测定脑组织水含量。结果:与模型组比较,补阳还五汤组、银杏叶片组PI3K、AKT蛋白表达显著升高(P〈0.05),AQP4在星胶细胞终足膜上的集中表达程度显著升高(P〈0.05),EB含量、脑组织含水量显著降低(P〈0.05)。结论:补阳还五汤对脑出血的保护作用机制可能与其能够激活PI3K/AKT信号通路,调控AQP4的"极性"表达,降低血脑屏障通透性,减轻脑水肿有关。
OBJECTIVE To explore effects of Buyanghuanwu Decoction on PI3K/AKT signaling pathway and AQP4 in cerebral hemorrhage rats,clarify mechanism of action and provide direction and target for therapy against cerebral hemorrhage caused by cerebral edema.METHODS SD rats were randomly divided into six groups:sham operation group,model group,Buyanghuanwu Decoction at low,moderate and high doses,Ginkgo biloba leaves.Rosenberg method was used to copy cerebral hemorrhage rat model.Expression of PI3 K and AKT was detected by immunohistochemistry.Expression of "polarity" of AQP4 was detected by immunofluorescence labeling method.Content of evans blue(EB)was determined by spectrophotometry.Water content of brain tissues was detected by wet and dry weight method.RESULTS Compared with model group,Buyanghuanwu Decoction group,ginkgo leaves group showed significantly increased expression of PI3 K and Akt proteins(P〈0.05),significantly increased concentrated expression of AQP4 in astrocyte(P〈0.05),significantly decreased EB and water contents in cerebral tissues(P〈0.05).CONCLUSION Protective effects of Buyanghuanwu Decoction against cerebral hemorrhage may be related with its capacity to activiate PI3K/AKT signal pathway,regulate expression of "polarity" of AQP4,reduce permeability of blood brain barrier,and alleviate brain edema.