补阳还五汤对急性脑缺血再灌注大鼠脑组织AKT和p-AKT蛋白表达的影响

目的:探讨补阳还五汤对急性脑缺血再灌注大鼠脑组织蛋白激酶B(AKT)和磷酸化蛋白激酶B(p-AKT)蛋白表达的影响,分析其对脑缺血再灌注损伤的保护机制。方法:将SPF级SD大鼠随机分为假手术组、模型组、氯吡格雷组、补阳还五汤高、低剂量组,每组20只。氯吡格雷组,补阳还五汤高,低剂量组分别按照动物体表面积换算,灌胃给予氯吡格雷6.8mg·kg-1·d-1,补阳还五汤26,6.5g·kg-1·d-1,其余各组给予蒸馏水。连续给药14d后,采用双侧颈总动脉阻断诱导大鼠急性脑缺血再灌注模型,比较各组病理组织学、脑组织皮层和海马CA1区AKT和p-AKT的表达水平。结果:HE染色结果提示,与模型组比较,补阳还五汤高,低剂量组能明显改善急性脑缺血再灌注大鼠脑组织损伤程度,随剂量增大而逐渐增强;免疫组化结果显示,各组的皮层和海马CA1区胞核和胞浆均可见AKT和p-AKT的阳性表达。与模型组比较,氯吡格雷组、补阳还五汤高剂量组AKT的阳性表达明显增加(P<0.05);与假手术组相比,模型组p-AKT的阳性表达明显降低(P<0.01);与模型组比较,氯吡格雷组、补阳还五汤高剂量组p-AKT阳性表达明显增加(P<0.05)。结论:补阳还五汤对急性脑缺血再灌注损伤模型大鼠的脑保护作用机制可能与上调PI3K/AKT信号通路中AKT,p-AKT的表达,促进AKT的磷酸化有关。

Influence of Buyang Huanwu Tang on Expressions of AKT and Phosphorylated-AKT in Acute Cerebral Ischemia-reperfusion Rat

Objective: To investigate the influence of Buyang Huanwu Tang (BHT)on the expressions of protein kinase B(AKT)and phosphorylated protein kinase B(p-AKT)in acute cerebral ischemia reperfusion rat. Method: The specific pathogen free SD rats were randomly divided into sham operation group, model group, positive control group and BHT(high, low dose) groups, each group contains 20 rats, Clopidogrel dose was 6.8 mg · kg^-1, and BHT high low dose was26, 6.5 g · kg^-1. The other groups were given distilled water. The acute cerebral ischemia-reperfusion models were established by bilateral carotid artery occlusion, using HE staining to observe Bcerebral pathological histology, using immunohistochemical method to detect brain cortex and hippocampus CA1 area AKT and p-AKT positive expression level. Result: BHT could significantly improve the degree of acute cerebral ischemia-reperfusion injury in rat brain tissue.HE Staining showed that the number of cortex neuron cells of acute cerebral ischemia-reperfusion rats decreased, cell arranged loosely and not regular, cell nucleus become atrophy, cells surrounding space became big obviously. The number of hippocampus CA1 area neurons decreased significantly, disordered arrangement of cells, and each dose group tissue damage degree were eased. Immunohistochemical results showed that each group of cortex and hippocampus CA1 area were seen AKT(Ab473)and p-AKT (ser473) positive expression. Compared with the sham operation group, model group AKT (Ab473) positive expression had no significant difference. Compared with model group, positive control group, BHT high dose group positive expression of AKT was obviously increased (P<0.05). Conclusion: The neuroprotective effect of Buyang Huanwu Tang against acute cerebral ischemia-reperfusion in rat may be associated with upregulation of AKT and p-AKT.