基于RBMECs-PC12细胞共培养体系的缺糖缺氧/复氧损伤模型探讨当归-川芎药对活性成分脑保护作用机制OA

Aim To explore the brain protective mechanism of the seven active ingredients(chloro-genic acid,ferulic acid,caffeic acid,butenylphtha-lein,senkyunolide H,senkyunolide A,ligustilide)in Danggui-Chuanxiong herb pair(GX).Methods An indirect co-culture method was used to construct a co-culture system of rat brain microvascular endothelial cells and adrenal pheochromocytoma cells(RBMECs-PC12).After 2,4,6,and 12 hours of glucose depri-vation and hypoxia,the system was reoxygenated for 24 hours.CCK-8 method was used to screen the opti-mal modeling time and administration concentration.The cells were divided into the control group,OGD/R group and low,medium and high dose groups of seven components.Flow cytometry was used to detect the apoptosis rate and ROS level in the co-culture system.RT-PCR was employed to detect the transcriptional lev-els of key genes of VEGF-AKT/NF-κB signaling path-way in the co-culture system.Results 12 hours of glucose deprivation and hypoxia in RBMECs-PC12 co-culture system was determined to be the best time for establishing the model.Compared with the OGD/R group,all the seven components could significantly re-duce the ROS level and the apoptosis rate of RBMECs-PC12 co-culture system.Chlorogenic acid,ferulic acid and caffeic acid significantly down-regulated the mRNA transcription levels of NF-κB p65 and MMP2.Chlorogenic acid and ferulic acid can also significantly down-regulate the mRNA transcription levels of Vegfr2 and Akt1.Senkyunolide H can up-regulate the mRNA transcription levels of Vegfr2 and Akt1.Conclusion The active components of Danggui-Chuanxiong herb pair can exert brain protection by reducing oxidative stress injury,reducing cell apoptosis rate,and inhibit-ing VEGF-AKT/NF-κB signaling pathway in RBMECs-PC12 co-culture system.