摘要
Objective:To investigate the protective effect of Cordyceps sinensis extract (CSE)on injury of primary cultured rat brain microvascular endothelial cells (rBMECs) induced by oxygen-glucose deprivation (OGD).Methods:We isolated and cultured primary rBMECs in order to establish an in vitro OGD model.Cellular activity was detected using a cell counting kit to determine the appropriate dosage.The rBMECs were divided into control,model,low-,mid-,and high-dose (5,10,20 μg.mL-1) CSE groups under OGD for 6 hours.CSE was dissolved in cell culture medium to the appropriate concentration,passed through a 0.22 μm sterile filter,and administered for 12 hours before and during OGD.Cellular morphology was observed under a microscope.Lactate dehydrogenase level in cultural supernatant,superoxide dismutase activity,and the content of nitric oxide and malondialdehyde in cells were tested by colorimetric methods.Levels of tumor necrosis factor-α and interleukin-1 beta in cells were determined by enzyme-linked immunosorbent assay.Results:After 12-hour administration of CSE at the concentration of 5,10,20 iμg.mL-1 before and during OGD,compared with the model group,the CSE groups obviously alleviated the damage of rBMECs induced by OGD,inhibited the apoptosis and the necrosis of the cells,and improved cellular morphology of rBMECs.Additionally,compared with the model group,CSE also restrained lactate dehydrogenase leakage in hypoxic cells (P <.01),significantly increased superoxide dismutase activity (P <.05),and reduced the levels of nitric oxide,malondialdehyde,tumor necrosis factor-α,and interleukin-1 beta (P <.05).Conclusion:C.sinensis extract plays a significant role in protecting injured primary cultured rBMECs induced by OGD.The mechanism may be related with the increase of cellular antioxidative capacity and anti-inflammatory effect.
基金
This study was supported by the National Natural Science Foundation of China(81403319 and 81603453)
the Beijing Excellent Talent Project(2014000020124G114).
