The Protective Effects of Juglanin in Cerebral Ischemia Reduce Blood–Brain Barrier Permeability via Inhibition of VEGF/VEGFR2 Signaling

Abstract

Introduction

Ischemic brain injury due to stroke or other pathologies is a major contributor to disability and mortality worldwide. Upon the occurrence of stroke, neuronal cells undergo apoptosis due to the deprivation of oxygen and nutrients and failure of the blood–brain barrier (BBB). In the moments immediately following a stroke, widespread perfusion resulting from hyperpermeability is accompanied by an acute inflammatory response, which induces neovascularization and often permanent neurological injury. Vascular endothelial growth factor (VEGF) and its receptor VEGF receptor 2 (VEGFR2) have been targeted to suppress cerebral ischemia. Recently, natural products including flavonoids, such as juglanin, have been receiving increasing attention for their impressive physiological effects.

Methods

Twenty mg/kg body weight juglanin was administrated for 3 weeks before inducing middle cerebral artery occlusion (MCAO) in mice. The animal brain infarction volume, neurological deficit score, blood–brain barrier permeability, and the expression of tight junction proteins were evaluated. Endothelial permeability and tight junction protein expression were also assessed in brain microvascular endothelial cells (HMBVECs) exposed to oxygen–glucose deprivation/reperfusion (OGD/R).

Results

Juglanin significantly reduced occlusion-induced infarct volume and improved neurological score by suppressing BBB hyperpermeability. Juglanin inhibited both the mRNA and protein expression of VEGF and VEGFR2 and restored the normal expression of occludin and zonula occludens-1 (ZO-1), two important tight junction proteins, in MCAO mice. Meanwhile, the results of in vitro experiments show that the protective effects of juglanin against increased BBB permeability and reduced tight junction functionality are dependent on the VEGF/VEGFR2 signaling pathway, as evidenced by the capacity of exogenous VEGF-A to abolish the effects of juglanin.

Conclusion

Our findings indicate a potent ability of juglanin to prevent neuronal injury resulting from cerebral ischemia by modulating the VEGF/VEGFR2 signaling pathway. Further research will help elucidate the exact mechanisms behind the protective effects of juglanin.

Keywords:

• cerebral ischemia
• stroke
• juglanin
• VEGF
• VEGFR2
• tight junction proteins
• blood–brain barrier
• BBB permeability

Highlights

1. Juglanin reduces infarct volume and improves neurological score;

2. Juglanin reduces blood–brain barrier permeability by increasing the expression of occludin and ZO-1;

3. The protective effects of juglanin are mediated through inhibition of VEGF/VEGFR2 signaling;

4. The protective effects of juglanin are evident both in vivo in MCAO mice and in vitro in HBMVECs.

Disclosure

The authors report no conflicts of interest in this work.