Study of brain protective mechanism of active ingredients in Danggui-Chuanxiong herb pair based on glucose deprivation hypoxia/reoxygenation injury model of RBMECs-PC12 cell coculture system

AimTo explore the brain protective mechanism of the seven active ingredients (chlorogenic acid, ferulic acid, caffeic acid, butenylphthalein, senkyunolide H, senkyunolide A, ligustilide) in Danggui-Chuanxiong herb pair (GX).MethodsAn 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 deprivation and hypoxia, the system was reoxygenated for 24 hours. CCK-8 method was used to screen the optimal 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 levels of key genes of VEGF-AKT/NF-κB signaling pathway in the co-culture system.Results12 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 reduce 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.ConclusionThe active components of Danggui-Chuanxiong herb pair can exert brain protection by reducing oxidative stress injury, reducing cell apoptosis rate, and inhibiting VEGF-AKT/NF-κB signaling pathway in RBMECs-PC12 co-culture system.