Mild folate deficiency induces a proatherosclerotic phenotype in endothelial cells.
Low folate/high homocysteine (Hcy) is an established risk marker for cardiovascular disease (CVD). Some in vivo studies suggest low folate may independently contribute to CVD. To study the effects of mild folate deficiency on endothelial function, we adapted the EA.hy 926 endothelial cell line to growth in medium containing 23 nM folic acid (LO cells) or 9 microM folic acid (HI cells). Folate derivatives were substantially depleted in LO cells relative to HI cells. No differences were seen in intracellular homocysteine, S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), the SAM:SAH ratio, or global DNA methylation, and there was no consistent difference in secreted homocysteine. A greater percentage of LO than HI cells were in S phase of the cell cycle; supplementation of LO cells with thymidine/hypoxanthine prevented this. LO cells were more elongated than HI cells and did not form tight monolayers. Stress fibers were very prominent in LO but not HI cells. Treatment of LO cells with rho kinase inhibitors abolished stress fibers and partially normalized cell shape. LO cell monolayers were more permeable than HI cell monolayers at confluence, and MCP-1 mRNA and protein expression was higher in LO than HI cells. Our results suggest that mild folate deficiency is proatherosclerotic.
Department of Pharmacology, University of Pennsylvania School of Medicine, 153 Johnson Pavilion, 3620 Hamilton Walk, Philadelphia, PA 19104-6084, USA.
Karen S
KS
Brown
Yuehua
Y
Huang
Zhi-Yong
ZY
Lu
Wenying
W
Jian
Ian A
IA
Blair
Alexander S
AS
Whitehead
D050197:Atherosclerosis / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D002455:Cell Division; D002460:Cell Line; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D004730:Endothelium, Vascular / Q000378:metabolism* / Q000473:pathology; D005494:Folic Acid Deficiency / Q000150:complications* / Q000378:metabolism / Q000473:pathology; D015870:Gene Expression; D006801:Humans; D010641:Phenotype; D012313:RNA / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012307:Risk Factors; D012435:S-Adenosylhomocysteine / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism
D018932:Chemokine CCL2; D012313:RNA; D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine