title: Tanshinone IIA protects the human blood-brain barrier model from leukocyte-associated hypoxia-reoxygenation injury.
abstract: To investigate the in vitro effect of tanshinone IIA on leukocyte-associated hypoxia-reoxygenation injury of human brain-blood barrier (BBB), we established the BBB model by culturing purified primary human brain microvascular endothelial cells (HBMVEC) to confluence on cell culture insert. BBB was identified by tight junction, transendothelial electrical resistance (TEER) and the permeability of BBB to horseradish peroxidase (HRP). The effect of tanshinone IIA on the permeability of BBB was tested at 2 h after hypoxia and 1h after reoxygenation with or without the supernatants of activated leukocytes. The effect of tanshinone IIA on leukocytes activation was analyzed by detection of MMP-9, cytokines and reactive oxygen species. The results showed that BBB formed by confluent HBMVECs had no cellular gap. Immunofluorescent staining for ZO-1 confirmed that the cells were connected by tight junction. Moreover, the BBB model had a higher TEER and a lower permeability for HRP than confluent HUVECs. The permeability of BBB for HRP was enhanced by hypoxia-reoxygenation and further greatly enhanced by adding the supernatants of activated leukocytes before reoxygenation. But such an effect was reversed by addition of tanshinone IIA before hypoxia. Moreover, tanshinone IIA could decrease the levels of MMP-9, TNF-α, IL-1α, IL-2, IFN-γ and reactive oxygen species in leukocytes. In conclusion, tanshinone IIA can protect BBB against leukocyte-associated hypoxia-reoxygenation injury by attenuating the activation of leukocytes and inhibiting the injury effects of leukocytic products. Tanshinone IIA may be a novel therapeutic agent for cerebral ischemia-reperfusion injury.
authors.affiliation: Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, PR China.
authors.forename: Wen-jian
authors.initials: WJ
authors.lastname: Zhang
authors.forename: Jie
authors.initials: J
authors.lastname: Feng
authors.forename: Ran
authors.initials: R
authors.lastname: Zhou
authors.forename: Li-ya
authors.initials: LY
authors.lastname: Ye
authors.forename: Hong-lin
authors.initials: HL
authors.lastname: Liu
authors.forename: Liang
authors.initials: L
authors.lastname: Peng
authors.forename: Jin-ning
authors.initials: JN
authors.lastname: Lou
authors.forename: Cheng-hui
authors.initials: CH
authors.lastname: Li
mesh_terms: D045784:Abietanes; D001812:Blood-Brain Barrier / Q000187:drug effects* / Q000293:injuries* / Q000378:metabolism / Q000473:pathology; D017077:Culture Media, Conditioned / Q000494:pharmacology; D017097:Electric Impedance; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D006801:Humans; D000860:Hypoxia / Q000378:metabolism* / Q000473:pathology; D007962:Leukocytes / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D010539:Permeability / Q000187:drug effects; D010616:Phenanthrenes / Q000494:pharmacology*; D015427:Reperfusion Injury / Q000517:prevention & control
chemical_list: D045784:Abietanes; D017077:Culture Media, Conditioned; D010616:Phenanthrenes; C021751:tanshinone; D010100:Oxygen