桃红四物汤对缺糖缺氧大鼠脑微血管内皮细胞VEGF、VEGFR-2、Akt基因及蛋白表达的作用机制研究

摘要 目的：探讨桃红四物汤对缺糖缺氧（OGD）大鼠脑微血管内皮细胞（rBMECs）血管内皮生长因子（VEGF）、血管内皮生长因子受体2（VEGFR-2）、蛋白激酶B（Akt）基因及蛋白表达的影响及其作用机制。方法：取rBMECs进行培养并随机分为正常组培养、模型组培养、桃红四物汤（0.4 mg/mL）组和桃红四物汤（0.8 mg/mL）组，除正常组外，其余三组采用氧糖剥夺实验构建OGD损伤模型。桃红四物汤组在造模前1 h按0.4 mg/mL、0.8 mg/mL浓度加入桃红四物汤，而正常组及模型组则加入等量体积生理盐水。实时定量聚合酶链反应（qRT-PCR）、蛋白免疫印迹试验检测VEGF、VEGFR-2、Akt基因及蛋白表达情况。结果：与正常组比较，模型组细胞凋亡率显著升高（P＜0.05）。与模型组比较，桃红四物汤（0.4 mg/mL）组和桃红四物汤（0.8 mg/mL）组均降低，且以桃红四物汤（0.8 mg/mL）组最为显著（P＜0.05）。与正常组比较，模型组rBMECs中Akt、VEGFR-2、VEGF相对mRNA和蛋白表达水平显著升高（P＜0.05）。与模型组比较，桃红四物汤（0.4 mg/mL）组和桃红四物汤（0.8 mg/mL）组显著降低，且以桃红四物汤（0.8 mg/mL）组最为显著（P＜0.05）。结论：桃红四物汤可能通过降低OGD条件下rBMECs中VEGF、VEGFR-2、Akt基因及蛋白表达水平，从而发挥抑制内皮细胞的凋亡，保持内皮细胞的完整性，这可能是桃红四物汤对脑保护的机制之一。

Study on the Mechanism of Taohong Siwu Decoction on the Expression of VEGF, VEGFR-2, Akt Gene and Protein in Brain Microvascular Endothelial Cells of Rats with Oxygen Glucose Deprivation

ABSTRACT Objective: To investigate the effect of Taohong Siwu decoction on the expression of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (VEGFR-2), protein kinase B (Akt) gene and protein in brain microvascular endothelial cells (rBMECs) of rats with oxygen glucose deprivation (OGD) and its mechanism. Methods: The rBMECs were cultured and randomly divided into normal group, model group, Taohong Siwu decoction (0.4 mg/mL) group and Taohong Siwu decoction (0.8 mg/mL) group. In addition to the normal group, the other three groups used oxygen glucose deprivation experiment to construct OGD injury model. Taohong Siwu decoction group was added with Taohong Siwu decoction at a concentration of 0.4 mg/mL and 0.8 mg/mL 1 h before modeling, while the normal group and the model group were added with the same volume of normal saline. Real-time quantitative polymerase chain reaction (qRT-PCR) and Western blotting were used to detect the expression of VEGF, VEGFR-2 and Akt genes and proteins. Results: Compared with the normal group, the apoptosis rate of the model group was significantly increased(P<0.05). Compared with the model group, the Taohong Siwu decoction (0.4 mg/mL) group and Taohong Siwu decoction (0.8 mg/mL) group decreased, and the Taohong Siwu decoction (0.8 mg/mL) group was the most significant(P<0.05). Compared with the normal group, the relative mRNA and protein expression levels of Akt, VEGFR-2 and VEGF in rBMECs of the model group were significantly increased(P<0.05). Compared with the model group, the Taohong Siwu decoction (0.4 mg/mL) group and Taohong Siwu decoction (0.8 mg/mL) group were significantly decreased, and the Taohong Siwu decoction (0.8 mg/mL) group was the most significant. Conclusion: Taohong Siwu decoction may inhibit the apoptosis of endothelial cells and maintain the integrity of endothelial cells by reducing the expression levels of VEGF, VEGFR-2 and Akt genes and proteins in rBMECs under OGD conditions,which may be one of the mechanisms of Taohong Siwu Decoction on brain protection.