Genomic changes in regenerated porcine coronary arterial endothelial cells.
OBJECTIVE
Genomic changes were defined in cultures of regenerated porcine coronary endothelial cells to explain the alterations that underlie their dysfunction.


RESULTS
Regeneration of the endothelium was triggered in vivo by endothelial balloon denudation. After 28 days, both left circumflex (native cells) and left anterior descending (regenerated cells) coronary arteries were dissected, their endothelial cells harvested, and primary cultures established. The basal cyclic GMP production was reduced in regenerated cells without significant reduction in the response to bradykinin and A23187. The mRNA expression levels in both native and regenerated cells were measured by microarray and RT-PCR. The comparison revealed genomic changes related to vasomotor control (cyclooxygenase-1, angiotensin II receptor), coagulation (F2 and TFPI), oxidative stress (Mn SOD, GPX3, and GSR), lipid metabolism (PLA2 and HPGD), and extracellular matrix (MMPs). A-FABP and MMP7 were induced by regeneration. RT-PCR revealed upregulation of A-FABP and downregulation of eNOS and TR. The differential gene expression profiles were confirmed at the protein level by Western blotting for eNOS, F2, Mn SOD, MMP7, and TR.


CONCLUSIONS
Cultures from regenerated coronary endothelial cells exhibit genomic changes explaining endothelial dysfunction and suggesting facilitation of coagulation, lipid peroxidation, and extracellular matrix remodeling.
Lee
Mary Y K
MY
Department of Pharmacology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 2F Laboratory Block, 21 Sassoon Road, Pokfulam, Hong Kong.
Tse
Hung-Fat
HF
Siu
Chung-Wah
CW
Zhu
Shu-Guang
SG
Man
Ricky Y K
RY
Vanhoutte
Paul M
PM
D000818:Animals; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000293:injuries; D006152:Cyclic GMP / Q000378:metabolism; D042783:Endothelial Cells / Q000502:physiology*; D005109:Extracellular Matrix / Q000378:metabolism; D050556:Fatty Acid-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D050356:Lipid Metabolism; D020783:Matrix Metalloproteinase 7 / Q000235:genetics / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress; D012333:RNA, Messenger / Q000378:metabolism; D012038:Regeneration / Q000235:genetics* / Q000502:physiology; D034421:Sus scrofa; D013880:Thioredoxin-Disulfide Reductase / Q000235:genetics* / Q000378:metabolism
D050556:Fatty Acid-Binding Proteins; D012333:RNA, Messenger; D052250:Nitric Oxide Synthase Type III; D013880:Thioredoxin-Disulfide Reductase; D020783:Matrix Metalloproteinase 7; D006152:Cyclic GMP