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  

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

作　　者：胡小伟 李琳[1] 龚荧荧 方燕[1] 杨琰[1] 许家栋[1] 储利胜[1] HU Xiaowei;LI Lin;GONG Yingying;FANG Yan;YANG Yan;XU Jiadong;CHU Lisheng(College of Basic Medical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China;The Second Clinical Medical College,Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学基础医学院,浙江杭州310053 [2]浙江中医药大学第二临床医学院,浙江杭州310053

出　　处：《浙江大学学报（医学版）》2022年第5期544-551,共8页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金(81073075);浙江省自然科学基金(LY22H280009);浙江中医药大学校级科研基金项目(2019ZY20,2021JKZC01)。

摘　　要：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。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)-1αand 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-1αand VEGF were up-regulated(all P<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-1αand VEGF through activation of PI3K-AKT signaling pathway.

关 键 词：补阳还五汤 脑微血管内皮细胞 氧糖剥夺再灌注 血管生成 PI3K-AKT通路 低氧诱导因子-1Α 血管内皮生长因子 

分 类 号：R285.5[医药卫生—中药学]