补阳还五汤对脑缺血再灌注损伤大鼠血小板Src,Akt和p38 MAPK蛋白的影响

目的:探讨补阳还五汤预处理对脑缺血再灌注损伤大鼠体内血小板活化信号通路中酪氨酸激酶(Src),蛋白激酶B(Akt)和p38丝裂原活化蛋白激酶(p38 MAPK)蛋白及其磷酸化表达的影响。方法:将SD大鼠随机分为正常组、假手术组、模型组、硫酸氢氯吡格雷组(7.81 mg·kg-1)和补阳还五汤高、中、低剂量组(29.69,14.84,7.42 g·kg~(-1))。连续灌胃ig给药14 d后,大脑中动脉线栓法复制脑缺血再灌注损伤模型,并神经功能缺损评分;氯化三苯基四氮唑(TTC)染色法测算脑梗死体积;蛋白质免疫印迹法(Western blot)检测血小板活化信号通路中Src,Akt和p38 MAPK蛋白及其磷酸化表达情况。结果:与模型组比较,补阳还五汤各组及硫酸氢氯吡格雷组能明显降低脑缺血再灌注损伤大鼠的神经功能缺损评分和脑梗死体积比(P0.05);补阳还五汤不同剂量明显降低Src,Akt和p38 MAPK磷酸化水平(P0.05);硫酸氢氯吡格雷明显降低Src和Akt磷酸化水平(P0.05);各组大鼠血小板Src,Akt和p38 MAPK总蛋白表达水平无统计学差异。结论:补阳还五汤预处理对脑缺血再灌注损伤大鼠的脑组织保护作用,一定程度上是发挥抗血小板活化的作用,且可能与抑制Src家族激酶,PI3K-Akt和p38 MAPK信号通路有关。

Effect of Buyang Huanwu Tang on Platelet Proteins Src, Akt and p38 MAPK of Cerebral Ischemia Reperfusion Injury Rat

Objective: To investigate the effect of pre-treatment with Buyang Huanwu Tang (BHT) on Src, Akt and p38 MAPK proteins and their phosphorylation expressions in vivo platelet activation signaling pathways of rats with cerebral ischemia-reperfusion (I/R) injury.Method: SD rats were randomly divided into blank control group, sham-operation group, model group, Clopidogrel Hydrogen Sulfate group (7.81 mg·kg^-1) and high-dose, medium-dose and low-dose BHT group (29.69, 14.84, 7.42 g·kg^-1). After oral administration with drugs for 14 d, the middle cerebral artery occlusion (MCAO) method was used to replicate the model of cerebral I/R injury, neurological deficit scores were measured. Besides, 2, 3, 5-Triphenyl tetrazolium chloride (TTC) staining method was used to calculate the volume of cerebral infarction; Western blot was used to measure platelet activation signaling pathways Src, Akt and p38 MAPK protein and their phosphorylation expressions. All the data were gathered after 14 days of continuous ig administration.Result: Compared with the model group, BHT at different doses and Clopidogrel Hydrogen reduced neurological deficit score and infarct volume percentage significantly in rats with cerebral I/R injury (P<0.05). BHT at different doses could significantly decrease Src, Akt and p38 MAPK phosphorylation levels (P<0.05). Clopidogrel Hydrogen dramatically reduced Src and Akt phosphorylation levels (P<0.05). Each group showed no statistical difference in total protein expression of Src, Akt and p38 MAPK.Conclusion: Pre-treatment with BHT had a cerebral protective effect on rats with cerebral I/R injury, which is partially attributed to its anti-platelet activation effects, and may be related with the inhibition of Src family kinases, PI3K-Akt and p38 MAPK signaling pathway.