Autophagy protects human brain microvascular endothelial cells against methylglyoxal-induced injuries, reproducible in a cerebral ischemic model in diabetic rats.

Abstract

Cerebral microvascular endothelial cells (ECs) are crucial for brain vascular repair and maintenance, but their physiological function may be impaired during ischemic stroke and diabetes. Methylglyoxal (MGO), a reactive dicarbonyl produced during glucose metabolism, could exacerbate ischemia-induced EC injury and dysfunction. We investigated the protective effect of autophagy on cultured human brain microvascular endothelial cells (HBMEC) that underwent MGO treatment. A further study was conducted to explore the underlying mechanisms of the protective effect. Autophagic activity was assessed by evaluating protein levels, using western blot. 3-methyladenine (3-MA), bafilomycin A1, ammonium chloride (AC), Beclin 1 siRNA, and chloroquine (CQ) were used to cause autophagy inhibition. Alarmar blue assay and lactate dehydrogenase release assay were used to evaluate cell viability. Streptozotocin was administered to induce type I diabetes in rats and post-permanent middle cerebral artery occlusion was performed to elicit cerebral ischemia. Blood-brain barrier permeability was also assessed. Our study found that MGO reduced HBMEC cell viability in a concentration- and time-dependent manner, and triggered the responsive autophagy activation. Autophagy inhibitors bafilomycin A1, AC, 3-MA, and BECN1 siRNA exacerbated MGO-induced HBMEC injury. FAK phosphorylation inhibitor PF573228 inhibited MGO-triggered autophagy and enhanced lactate dehydrogenase release. Meanwhile, similar autophagy activation in brain vascular ECs was observed during permanent middle cerebral artery occlusion-induced cerebral ischemia in diabetic rats, while chloroquine-induced autophagy inhibition enhanced blood-brain barrier permeability. Taken together, our study indicates that autophagy triggered by MGO defends HBMEC against injuries.

• Authors: Lili Fang, Xue Li, Yinbo Zhong, Jing Yu, Lina Yu, Haibin Dai, Min Yan
• Journal: Journal of neurochemistry (J Neurochem) Vol. 135 Issue 2 Pg. 431-40 (Oct 2015) ISSN: 1471-4159 [Electronic] England
• PMID: 26251121 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2015 International Society for Neurochemistry.

Chemical References

• Pyruvaldehyde
• Focal Adhesion Kinase 1

Topics

• Animals
• Autophagy (drug effects)
• Blood-Brain Barrier (drug effects)
• Brain Ischemia (chemically induced, drug therapy)
• Capillaries (cytology, drug effects)
• Cell Line
• Cell Survival (drug effects)
• Diabetes Mellitus, Experimental (pathology)
• Diabetes Mellitus, Type 1 (chemically induced, pathology)
• Diabetic Angiopathies (pathology)
• Endothelial Cells (drug effects)
• Focal Adhesion Kinase 1 (metabolism)
• Humans
• Male
• Middle Cerebral Artery (pathology)
• Pyruvaldehyde (toxicity)
• RNA Interference (drug effects)
• Rats
• Rats, Sprague-Dawley