Effect of Withanolide A on 7-Ketocholesterol Induced Cytotoxicity in hCMEC/D3 Brain Endothelial Cells

• Published in: Cells (Basel, Switzerland) Vol. 11; no. 3; p. 457
• Main Authors: Soh, Sandra, Ong, Wei-Yi
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 28.01.2022; MDPI
• Subjects: 7-ketocholesterol; Antioxidants; Apoptosis; Arteriosclerosis; Blood coagulation; blood-brain-barrier; Brain; brain endothelial cells; Cardiovascular disease; Cell culture; Cell cycle; Cell viability; Cholesterol; Clotting; Cyclooxygenase 2 - genetics; Cyclooxygenase-2; Cytotoxicity; Diabetes mellitus; Endothelial Cells; Enzymatic activity; Experiments; Ginseng; Glucocorticoids; Humans; Hypercholesterolemia; IL-1β; Inflammation; Interleukin 6; Interleukin 8; Ketocholesterols - pharmacology; Mifepristone; Nitric-oxide synthase; Oxidation; Plaques; Receptors, Glucocorticoid; Thrombin; Thrombin - pharmacology; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Von Willebrand factor; withanolide A; Withanolides; Ann Arbor Michigan; United States > US; blood-brain-barrier; 7-ketocholesterol; cerebral ischemia; vascular dementia; brain endothelial cells; diabetes mellitus; withanolide A; hypercholesterolemia
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells11030457

Withanolide A is a naturally occurring phytochemical that is found in Ashwagandha ( , fam. Solanaceae) or Indian Ginseng. In the current study, we elucidated the effect of withanolide A on 7-ketocholesterol (7KC) induced injury in hCMEC/D3 human brain endothelial cells. 7KC is a cholesterol oxidation product or oxysterol that is present in atherosclerotic plaques and is elevated in the plasma of patients with hypercholesterolemia and/or diabetes mellitus. Results showed that withanolide A significantly reduced the effects of 7KC, which include loss of endothelial cell viability, increase in expression of pro-inflammatory genes-IL-1β, IL-6, IL-8, TNF-α, cyclooxygenase-2 (COX-2), increased COX-2 enzyme activity, increased ROS formation, increased expression of inducible nitric oxide synthase and genes associated with blood clotting, including Factor 2/thrombin, Factor 8, von Willebrand factor, and thromboxane A synthase, and increased human thrombin enzyme activity. Some of the above effects of withanolide A on 7KC were reduced in the presence of the glucocorticoid receptor antagonist, mifepristone (RU486). These findings suggest that the glucocorticoid receptor could play a role in the cytoprotective, antioxidant, and anti-clotting effects of withanolide A against 7KC. Further studies are necessary to elucidate the detailed mechanisms of action of withanolide A against oxysterol-induced injury.