四君子汤加减对脑缺血/再灌注损伤大鼠脑组织ERK1/2,Akt,Bax表达的影响

Effect of Modified Si Junzitang on Expression of ERK1/2,Akt,Bax in Rats with Cerebral Ischemia/Reperfusion Injury

目的：观察四君子汤加减对脑缺血/再灌注损伤大鼠细胞外调节蛋白激酶1/2（ERK1/2）,蛋白激酶B（Akt）,细胞淋巴瘤/白血病-2相关X蛋白（Bax）的影响,探讨其保护神经细胞可能作用机制。方法：将55只雄性SD大鼠随机分为假手术组、模型组、依达拉奉组、四君子汤加减低、中、高剂量组（3.69,7.38,14.76 g·kg-1）。采用线栓法制备大脑中动脉阻塞（MCAO）模型,治疗7 d后处死,取大鼠缺血侧脑组织,运用蛋白免疫印迹法（Western blot）检测磷酸化ERK1/2（p-ERK1/2）,磷酸化Akt（p-Akt）,Bax蛋白含量。应用实时荧光定量聚合酶链式反应法（Real-time PCR）测定脑缺血再灌注后脑组织内ERK1/2,Akt,Bax mRNA的表达变化。结果：与假手术组比较,模型组p-ERK1/2,p-Akt蛋白及mRNA表达显著下调（P〈0.01）;Bax蛋白及mRNA表达显著上调（P〈0.01）;与模型组比较,四君子汤加减低、中、高剂量组p-ERK1/2,p-Akt蛋白及mRNA表达明显上调,Bax蛋白及mRNA表达明显下调,差异具有统计学意义（P〈0.01）。结论：四君子汤加减可能通过上调p-ERK1/2,p-Akt,下调Bax蛋白表达发挥脑保护作用。

Objective： To observe the effect of modefied Si Junzitang on the expression of extracellular signal-regulated kinase1/2（ ERK1/2）,protein kinase B（ Akt） and B cell lymphoma/lewkmia-2 assaciated X protein（ Bax） in rats with cerebral ischemia/reperfusion injury,and to explore its possible protective mechanism on nerve cells. Method： Totally 55 male SD rats were randomly divided into sham operation group,model group,edaravone group,modefied Si Junzitang low-dose group,middle-dose group,and high-dose group（ 3. 69,7. 38,14. 76 g·kg-1）. The middle cerebral artery occlusion（ MCAO） model was induced by suture method,after 7 d treatment,the rats were sacrificed and ischemic side brain tissues were taken. Phosphorylation of ERK1/2（ pERK1/2）,p-Akt and p-Bax protein contents were detected by Western blot; the mRNA expression levels of ERK1/2,Akt and Bax were detected by real time fluorescence quantitative polymerase chain reaction（ Real-time PCR）. Result： As compared with sham operation group,the p-ERK1/2,p-Akt and p-Bax protein and mRNA expression levels were significantly reduced in model group（ P 〈0. 01）. As compared with the model group,the protein and mRNA expression levels of p-ERK1/2 and p-Akt were up-regulated,the expression of protein and mRNA of Bax were significantly reduced in low-dosed group,middle-dosed group,high-dosed group,with statistically significant difference（ P 〈0. 01）. Conclusion： Modefied Si Junzitang can protects brain with cerebral ischemia/reperfusion injury by up-regulating the expression of p-ERK1/2 and p-Akt. reducing the expression of Bax.