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Updated: Dec 18, 2025
Analyzing the Permeability of the Blood-Brain Barrier by Microbial Traversal through Microvascular Endothelial Cells
Published on: February 14, 2020
GKT136901 protects primary human brain microvascular endothelial cells against methamphetamine-induced blood-brain
Jong Su Hwang1, Eun-Hye Cha1, Eunyoung Ha1
1Department of Biochemistry, School of Medicine, Keimyung University, Daegu 42601, Republic of Korea.
Aims:
Methamphetamine (METH) is an abused psychostimulant causing public health concern worldwide. While most studies have focused on the neurotoxic effects of METH, METH-induced cerebrovascular dysfunction has recently drawn attention as an important facet of METH-related pathophysiology. In this study, we investigated the protective role of GKT136901, a NOX1/4 inhibitor, against METH-induced blood-brain barrier (BBB) dysfunction.
Main Methods:
Primary human brain microvascular endothelial cells (HBMECs) were used as an in vitro BBB model. HBMECs were treated with GKT136901, followed by METH exposure for 24 h. The generation of reactive oxidative species (ROS) was measured using 2',7'-dichlorofluorescin diacetate (DCF-DA) staining. To examine the BBB function, paracellular permeability of HBMEC monolayer was measured using FITC-labeled dextran. To evaluate structural properties of BBB in HBMECs, tight junction (TJ), adherent junction (AJ), and cytoskeletal proteins were stained and analyzed by confocal microscopy.
Key Findings:
METH treatment rapidly increased ROS generation in HBMECs but GKT136901 treatment inhibited METH-induced ROS generation. Although METH increased the permeability of HBMEC monolayer, this effect was abolished upon GKT136901 treatment. Following METH exposure, the proteins Zonula occludens-1 (ZO-1) and vascular endothelial cadherin (VE-cadherin) were translocalized from the cell membrane to the cytoplasm, thereby destroying intercellular tight junction (TJ) and adherent junction (AJ) structures, which were ameliorated upon GKT136901 treatment. METH exposure altered the cellular morphology of HBMECs and induced stress fiber formation. However, GKT136901 prevented METH-induced morphological and cytoskeletal changes in HBMECs.
Significance:
These results suggest that GKT136901 prevents METH-induced BBB dysfunction in HBMECs through the inhibition of ROS generation.
Insights
This study shows GKT136901 protects against methamphetamine-induced blood-brain barrier (BBB) dysfunction. The NOX1/4 inhibitor reduced reactive oxidative species (ROS) and preserved BBB integrity in human brain cells.
Area of Science:
- Neuroscience
- Pharmacology
- Cell Biology
Background:
- Methamphetamine (METH) abuse poses a global health risk, with increasing focus on its cerebrovascular effects.
- METH-induced blood-brain barrier (BBB) dysfunction is a critical but understudied aspect of its pathophysiology.
Purpose of the Study:
- To investigate the protective potential of GKT136901, a novel NOX1/4 inhibitor, against METH-induced BBB dysfunction.
- To elucidate the mechanisms underlying GKT136901's protective effects on the BBB.
Main Methods:
- Utilized primary human brain microvascular endothelial cells (HBMECs) as an in vitro BBB model.
- Assessed reactive oxidative species (ROS) generation, paracellular permeability, and tight junction (TJ) and adherent junction (AJ) protein integrity.
- Analyzed cellular morphology and cytoskeletal changes using confocal microscopy.
Main Results:
- METH significantly increased ROS generation and BBB permeability in HBMECs.
- GKT136901 treatment inhibited METH-induced ROS production and prevented the increase in HBMEC monolayer permeability.
- GKT136901 preserved the localization of ZO-1 and VE-cadherin, maintaining TJ and AJ structures and preventing METH-induced morphological and cytoskeletal damage.
Conclusions:
- GKT136901 demonstrates significant protective effects against METH-induced BBB dysfunction in vitro.
- Inhibition of ROS generation is a key mechanism by which GKT136901 confers BBB protection.
- GKT136901 represents a potential therapeutic strategy for mitigating METH-related cerebrovascular damage.
