Both lisinopril and verapamil reduced platelet-derived growth factor-A chain mRNA levels in human saphenous vein endothelial cells stimulated by thrombin.

Background: Both angiotensin-converting enzyme inhibitors and calcium channel blockers decrease postinjury intimal thickening in vivo, but their mechanisms of inhibitory action are unclear. Expression of the gene for platelet-derived growth factor (PDGF), a smooth-muscle mitogen, in endothelial cells (ECs) after vessel injury has been postulated to cause intimal thickening. In this study, we tested whether lisinopril, an angiotensin-converting enzyme inhibitor, or verapamil, a calcium channel blocker, would suppress the PDGF gene expression in stimulated human saphenous vein ECs.

Heparin and dibutyryl cAMP modulate gene expression in stimulated human saphenous vein smooth muscle cells.

Increased expression of basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF) A chain, and tissue plasminogen activator (tPA) by smooth muscle cells (SMC) has been postulated to mediate the progression of intimal hyperplasia. We tested whether heparin would suppress the expression of these genes in stimulated human saphenous vein SMC. Quiescent cultured human saphenous vein SMC were stimulated for 4 h with heat-inactivated fetal bovine serum (10% by vol) in the presence or absence of heparin (1 to 250 micrograms/ml).

Construction of overlapping oligonucleotide templates for the production of cRNA standards for quantitative reverse transcription polymerase chain reaction.

The reverse transcriptase polymerase chain reaction (PCR) is a technique for the study of gene expression that requires far less RNA for analysis than Northern blots. The inclusion of cRNA standards in the initial reverse transcription step is a way to control for the tube-to-tube variation often inherent in the technique and to permit quantitation of the starting amount of the native mRNA being analyzed. We describe a method using overlapping oligonucleotides to produce templates for the production of cRNA standards for up to three different mRNA species.

Hantaan virus infection of human endothelial cells.

The primary pathophysiologic finding of the viral disease known as Korean hemorrhagic fever, the etiological agent of which is Hantaan virus (HTV), is vascular instability. To investigate whether HTV was able to infect cells derived from human vascular tissue and alter their behavior, we infected in vitro primary adult human endothelial cells from saphenous veins (HSVEC). We were able to detect the presence of viral antigens in infected cells both by immunofluorescence and by Western blot (immunoblot) analysis as early as day 1 postinfection.