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Updated: May 8, 2026
Oxygen-Glucose Deprivation and Reoxygenation as an In Vitro Ischemia-Reperfusion Injury Model for Studying Blood-Brain Barrier Dysfunction
Published on: May 7, 2015
Modulation of P-glycoprotein in rat brain microvessel endothelial cells under oxygen glucose deprivation
Bian-Sheng Ji1, Juan Cen, Ling He
1Key Laboratory of Natural Medicine and Immune Engineering, Henan University, Kaifeng, China.
Objectives:
To investigate modulation of P-glycoprotein (P-gp) in rat brain microvessel endothelial cells (rBMECs) under oxygen glucose deprivation (OGD).
Methods:
The coculture of rBMECs and astrocytes was established to investigate the time course of P-gp, tumour necrosis factor-α (TNF-α), endothelin-1 (ET-1), nitric oxide synthase (NOS) and protein kinase C (PKC) expression in the rBMECs as well as rhodamine 123 (Rh123) transendothelial transfer under OGD using Western blot and HPLC, respectively. The influence of pharmacological tools including H398, JKC-301, RES-701-1, L-NMMA, BIM and SN50 on the P-gp expression as well as Rh123 transendothelial transfer was evaluated at 3 h time point of OGD.
Key Findings:
Elevated P-gp, TNF-α, ET-1, NOS and PKC expression in the rBMECs, as well as increased P-gp efflux activity were observed after 2 h or more time of OGD. Incubation of H398 and other pharmacological tools downregulated P-gp expression and functional activity in the rBMECs at 3 h time point of OGD.
Conclusions:
This report suggested that TNF-α, ET-1, NOS and PKC may mediate upregulation of P-gp in the rBMECs under OGD, which may be worthy of being referenced for the investigation of P-gp at the blood-brain barrier in the early period of stroke.
Insights
Oxygen glucose deprivation (OGD) upregulates P-glycoprotein (P-gp) in rat brain microvessel endothelial cells. Pharmacological agents downregulate this P-gp expression and activity, offering potential therapeutic insights for stroke.
Area of Science:
- Neuroscience
- Pharmacology
- Cell Biology
Background:
- The blood-brain barrier (BBB) protects the brain but can be compromised during stroke.
- P-glycoprotein (P-gp) plays a crucial role in regulating the passage of substances across the BBB.
- Understanding P-gp modulation during ischemic conditions is vital for stroke research.
Purpose of the Study:
- To investigate the modulation of P-glycoprotein (P-gp) in rat brain microvessel endothelial cells (rBMECs) under oxygen-glucose deprivation (OGD).
- To explore the role of specific signaling molecules and pharmacological agents in P-gp regulation during OGD.
Main Methods:
- Establishment of a co-culture model of rBMECs and astrocytes.
- Time-course analysis of P-gp, TNF-α, ET-1, NOS, and PKC expression using Western blot.
- Assessment of rhodamine 123 (Rh123) transendothelial transfer via HPLC.
- Evaluation of pharmacological agents' effects on P-gp expression and function at 3 hours of OGD.
Main Results:
- OGD significantly elevated P-gp, TNF-α, ET-1, NOS, and PKC expression in rBMECs after 2 hours or more.
- Increased P-gp efflux activity was observed under OGD conditions.
- Pharmacological agents, including H398, effectively downregulated P-gp expression and functional activity at 3 hours of OGD.
Conclusions:
- TNF-α, ET-1, NOS, and PKC signaling pathways may mediate the upregulation of P-gp in rBMECs during OGD.
- These findings provide a basis for investigating P-gp modulation at the BBB in the early stages of stroke.
- Targeting P-gp could be a potential therapeutic strategy for stroke management.
