Cyclic nucleotides and production of prostanoids in human varicose veins.
OBJECTIVE
Experiments were designed to determine the production of prostacyclin and thromboxane and the activation of cyclic nucleotides in human varicose and nonvaricose veins and to determine whether these second messenger pathways were differentially activated by the venotropic extract of Ruscus aculeatus.


METHODS
The experiments were designed to characterize the activity of cyclic nucleotides and the production of prostaglandins in human varicose and nonvaricose veins. Segments of the greater saphenous veins and the adjacent tributaries were obtained from patients who underwent vein stripping and excision of primary varicose veins. The saphenous veins from the patients who underwent peripheral arterial bypass grafting were used as controls. The segments of veins were incubated in Krebs-Ringer bicarbonate solution in the presence of venotropic extract of Ruscus aculeatus (10(-3) g/mL) or in water-miscible organic solvent (dimethyl sulfoxide, 10(-3) g/mL), for 1, 5, and 10 minutes at 37 degrees C. The nonspecific phosphodiesterase inhibitor (3-isobutyl-1-methylxanthine, 10(-4) g/mL) was used to block cyclic nucleotide degradation in some samples. Tissue and media samples were collected. Tissue concentrations of both cyclic adenosine monophosphate and cyclic guanosine monophosphate (cAMP and cGMP, respectively) and media concentrations of 6-ketoprostaglandin-F(1)(alpha) (the stable metabolite of prostacyclin) and thromboxane B(2) (the stable metabolite of thromboxane A(2)) were measured by means of radioimmunoassay. Cyclooxygenase 2 was measured with Western blot analysis.


RESULTS
The varicose veins showed greater levels of cAMP but not of cGMP at all time points as compared with the control veins. Prostanoid production was not significantly altered in the varicose veins. Stimulation with Ruscus aculeatus increased the cAMP concentration in the varicose veins but did not affect the cGMP levels. The ratio between 6-ketoprostaglandin-F(1)(alpha) and thromboxane B(2) was two-fold greater in the varicose veins as compared with the control veins. In the presence of the extract, the ratio of 6-ketoprostaglandin-F(1)(alpha) and thromboxane B(2) was identical in both types of veins. Cyclooxygenase 2 was not present in either the control or the varicose veins.


CONCLUSIONS
These results suggest that cAMP levels are elevated in varicose veins and that they can be altered with drug treatment in varicose veins. This chemical pathway may be considered as a modulatory target to affect contraction with venotropic drugs.
Nemcova
S
S
Division of Vascular Surgery, Department of Surgery, Mayo Clinic and Foundation, Rochester, Minnesota, USA.
Gloviczki
P
P
Rud
K S
KS
Miller
V M
VM
D015121:6-Ketoprostaglandin F1 alpha / Q000096:biosynthesis*; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000242:Cyclic AMP / Q000378:metabolism*; D006152:Cyclic GMP / Q000378:metabolism; D051546:Cyclooxygenase 2; D005260:Female; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis; D008297:Male; D008565:Membrane Proteins; D008875:Middle Aged; D010544:Peroxidases / Q000096:biosynthesis; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis; D012501:Saphenous Vein / Q000378:metabolism; D015290:Second Messenger Systems; D013929:Thromboxane B2 / Q000096:biosynthesis*; D014648:Varicose Veins / Q000378:metabolism*
D007527:Isoenzymes; D008565:Membrane Proteins; D013929:Thromboxane B2; D015121:6-Ketoprostaglandin F1 alpha; D000242:Cyclic AMP; D010544:Peroxidases; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; D006152:Cyclic GMP