cGMP accumulation and gene expression of soluble guanylate cyclase in human vascular tissue.
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.
Papapetropoulos
A
A
Department of Pharmacology and Toxicology, Medical College of Georgia, Augusta 30912, USA.
Cziraki
A
A
Rubin
J W
JW
Stone
C D
CD
Catravas
J D
JD
D000818:Animals; D001011:Aorta / Q000166:cytology; D009320:Atrial Natriuretic Factor / Q000494:pharmacology; D001483:Base Sequence; D002417:Cattle; D002642:Chick Embryo; D006152:Cyclic GMP / Q000378:metabolism*; D004285:Dogs; D004730:Endothelium, Vascular / Q000187:drug effects / Q000201:enzymology*; D015870:Gene Expression / Q000502:physiology; D006162:Guanylate Cyclase / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000201:enzymology*; D009599:Nitroprusside / Q000494:pharmacology; D010446:Peptide Fragments; D011651:Pulmonary Artery / Q000166:cytology; D011667:Pulmonary Veins / Q000166:cytology; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D051381:Rats; D012501:Saphenous Vein / Q000166:cytology; D012995:Solubility; D013552:Swine; D014665:Vasodilator Agents / Q000494:pharmacology
D007527:Isoenzymes; D010446:Peptide Fragments; D012333:RNA, Messenger; D014665:Vasodilator Agents; D009599:Nitroprusside; D009320:Atrial Natriuretic Factor; C062524:atrial natriuretic factor prohormone (103-125); D006162:Guanylate Cyclase; D006152:Cyclic GMP