补阳还五汤激活PI3K-AKT通路促进氧糖剥夺再灌注损伤的脑微血管内皮细胞血管生成

目的：探讨补阳还五汤促进氧糖剥夺再灌注（OGD/R）损伤的大鼠脑微血管内皮细胞（RBMEC）血管生成的机制。方法：RBMEC经补阳还五汤含药血清孵育24?h后，构建OGD/R细胞损伤模型。将细胞随机分为正常对照组、模型对照组、补阳还五汤组（给予补阳还五汤含药血清）和LY294002组给予补阳还五汤含药血清前使用磷脂酰肌醇3激酶（PI3K）抑制剂LY294002预处理1?h]。采用CCK-8法、细胞划痕实验和Transwell迁移实验、管腔形成实验分别检测细胞增殖、迁移和形成管腔能力。蛋白质印迹法检测PI3K、蛋白激酶B（AKT）、低氧诱导因子（HIF）-1α及血管内皮生长因子（VEGF）等蛋白的表达。结果：与模型对照组比较，补阳还五汤组RBMEC存活率、迁移能力及管腔形成能力显著增强（均P<0.01），磷酸化PI3K、磷酸化AKT、HIF-1α和VEGF蛋白表达增加（均P<0.05），而LY294002可阻断上述效应。结论：补阳还五汤含药血清可促进OGD/R损伤的RBMEC血管生成，其机制可能与其激活PI3K-AKT信号通路上调HIF-1α和VEGF表达有关。

Buyang Huanwu decoction promotes angiogenesis of rat brain microvascular endothelial cells after oxygen-glucose deprivation reperfusion injury via activation of PI3K-AKT signaling pathway

Objective: To investigate the effect and mechanism of Buyang Huanwu decoction (BYHWD) on angiogenesis of rat brain microvascular endothelial cells (RBMECs) after oxygen-glucose deprivation reperfusion (OGD/R) injury. Methods: RBMECs were pretreated with BYHWD containing serum 24?h before OGD/R injury was induced. Cells were randomly divided into blank control group, model control group, BYHWD group (provided BYHWD containing serum) and LY294002 group treated with phosphoinositide 3-kinase (PI3K) inhibitor LY294002 for 1?h before provided BYHWD containing serum]. The cell viability, migration and tube formation abilities of RBMECs were detected by CCK-8, scratch wound healing, Transwell migration and tube formation assays, respectively. The protein expression levels of PI3K, p-PI3K, protein kinase B (AKT), p-AKT, hypoxia-inducible factor (HIF)-1α and vascular endothelial growth factor (VEGF) were determined by Western blotting. Results: Compared with model control group, cell viability, migration and tube formation abilities of RBMECs were significantly improved in BYHWD group (all P<0.01), the protein expression levels of p-PI3K, p-AKT, HIF-1α and VEGF were up-regulated (allP<0.05); while above effects were blocked by LY294002.Conclusion: BYHWD can promote angiogenesis of RBMECs after OGD/R injury, which may be related to the increased protein expression of HIF-1α and VEGF through activation of PI3K-AKT signaling pathway.