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Gene expression of soluble guanylate cyclase (sGC) and cGMP accumulation in response to sodium nitroprusside (SNP) were studied in cultured human vascular cells and freshly harvested vascular tissue. As revealed by reverse transcriptase-polymerase chain reaction, cultured smooth muscle and endothelial cells, as well as freshly isolated human vascular tissue, express mRNA for the alpha 3 and beta 3 subunits but not for the alpha 2 and beta 3 subunits is evident even in the absence of increased cGMP accumulation in response to SNP. cGMP accumulation in human cells cultured from different vascular beds typically increased two- to five-fold (maximum of 11.4-fold) over baseline following stimulation with 100 microM SNP. Bovine, murine, canine, and avian vascular smooth muscle cells accumulated similar or lower amounts of cGMP than human cells, whereas porcine, rat, and rabbit smooth muscle cells accumulated greater amounts of cGMP. In freshly harvested human vessels, cGMP accumulation in response to SNP was found to increase fifteen-fold over baseline. In contrast to the SNP-induced cGMP accumulation, cGMP levels in response to particulate guanylate cyclase activator atriopeptin II were equal to or greater in cultured human cells than in fresh human vascular tissue. We conclude that human vascular cells (fresh and cultured) express the mRNA for both a large (alpha 3) and a small (beta 3) sGC subunit and that fresh human cells are more sensitive to SNP stimulation.

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Methylene blue is occasionally applied to the adventitia of blood vessels during coronary artery bypass and other vascular procedures to assist in the orientation of the vessel. Inherent in this method is the assumption that extravascular application of methylene blue is innocuous with regard to vascular function. In the first part of this study, the in vitro vascular reactivity of methylene blue-labeled saphenous veins was compared with that of veins that were not marked with methylene blue. The vasoactive agents tested were designed to examine multiple pathways. They included potassium chloride, prostaglandin F2 alpha, phenylephrine, serotonin, angiotensin II, BHT-933 (alpha 2-adrenergic agonist), sodium nitroprusside, acetylcholine, isoproterenol, and verapamil. Compared with unmarked veins, those marked with methylene blue demonstrated a significant impairment of both vasoconstrictor and vasodilator function. These observations were made on a relatively small number of patients and could therefore be attributed to inherent differences between patients or surgical procedures. In the second part of this study, these variables were eliminated by dividing a single vein from one patient into three segments for a 45-minute exposure to external only methylene blue, internal and external methylene blue, or no methylene blue. The segments were then evaluated for vasoreactivity in vitro. Externally applied methylene blue reduced vasoconstriction regardless of the agonist. Further, both endothelium-dependent and -independent vasodilation was diminished by external methylene blue exposure. In veins exposed to methylene blue both internally and externally the results were similar but the magnitude of impairment greater. It is concluded that surgical marking of blood vessels with methylene blue has the potential to adversely affect vascular reactivity and therefore the use of alternative dyes should be considered.