Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1995 Sep;96(3):1449–1454. doi: 10.1172/JCI118181

Cyclic strain upregulates nitric oxide synthase in cultured bovine aortic endothelial cells.

M A Awolesi 1, W C Sessa 1, B E Sumpio 1
PMCID: PMC185768  PMID: 7544806

Abstract

In vivo, endothelial cells (EC) are subjected to hemodynamic forces which may influence the production of nitric oxide. This study was designed to examine the effect of cyclic strain on the expression of endothelial nitric oxide synthase (eNOS) in cultured bovine aortic EC. EC were grown on flexible membranes which were subjected to deformation at 60 cycles/min with -5 or -20 kPa of vacuum. This results in an average strain of 6 and 10%, respectively, which is transmitted to the attached cells. Northern blot analysis of total cytosolic RNA demonstrated an increase in eNOS gene expression with both strain regimens but the increase with 10% average strain was greater than that at 6%. Nuclear runoff transcription assays confirmed the induction of eNOS transcripts. Western blot analysis showed an increase in eNOS level after 24 h of cyclic 10% average strain compared with controls or 6% average strain. Immunohistochemical staining of EC for eNOS was increased in the high strain periphery (7-24% strain) of membranes deformed with -20 kPa vacuum. These results demonstrate that cyclic strain upregulates the expression of eNOS transcripts and protein levels in bovine aortic EC thus emphasizing the importance of hemodynamic forces in the regulation of eNOS in vivo.

Full text

PDF
1449

Images in this article

1451Image
on p.1451
1451Image
on p.1451
1451Image
on p.1451
1451Image
on p.1451
1451Image
on p.1451
1451Image
on p.1451
1451Image
on p.1451
1452Image
on p.1452
1452Image
on p.1452

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ando J., Ohtsuka A., Korenaga R., Sakuma I., Kamiya A. Flow-induced calcium transients and release of endothelium-derived relaxing factor in cultured vascular endothelial cells. Front Med Biol Eng. 1993;5(1):17–21. [PubMed] [Google Scholar]
  2. Awolesi M. A., Widmann M. D., Sessa W. C., Sumpio B. E. Cyclic strain increases endothelial nitric oxide synthase activity. Surgery. 1994 Aug;116(2):439–445. [PubMed] [Google Scholar]
  3. Banes A. J., Gilbert J., Taylor D., Monbureau O. A new vacuum-operated stress-providing instrument that applies static or variable duration cyclic tension or compression to cells in vitro. J Cell Sci. 1985 Apr;75:35–42. doi: 10.1242/jcs.75.1.35. [DOI] [PubMed] [Google Scholar]
  4. Bergel D. H. The static elastic properties of the arterial wall. J Physiol. 1961 May;156(3):445–457. doi: 10.1113/jphysiol.1961.sp006686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Delp M. D., McAllister R. M., Laughlin M. H. Exercise training alters endothelium-dependent vasoreactivity of rat abdominal aorta. J Appl Physiol (1985) 1993 Sep;75(3):1354–1363. doi: 10.1152/jappl.1993.75.3.1354. [DOI] [PubMed] [Google Scholar]
  6. Diamond S. L., Eskin S. G., McIntire L. V. Fluid flow stimulates tissue plasminogen activator secretion by cultured human endothelial cells. Science. 1989 Mar 17;243(4897):1483–1485. doi: 10.1126/science.2467379. [DOI] [PubMed] [Google Scholar]
  7. Ehsani A. A., Heath G. W., Hagberg J. M., Sobel B. E., Holloszy J. O. Effects of 12 months of intense exercise training on ischemic ST-segment depression in patients with coronary artery disease. Circulation. 1981 Dec;64(6):1116–1124. doi: 10.1161/01.cir.64.6.1116. [DOI] [PubMed] [Google Scholar]
  8. Frangos J. A., Eskin S. G., McIntire L. V., Ives C. L. Flow effects on prostacyclin production by cultured human endothelial cells. Science. 1985 Mar 22;227(4693):1477–1479. doi: 10.1126/science.3883488. [DOI] [PubMed] [Google Scholar]
  9. Graier W. F., Groschner K., Schmidt K., Kukovetz W. R. Increases in endothelial cyclic AMP levels amplify agonist-induced formation of endothelium-derived relaxing factor (EDRF). Biochem J. 1992 Dec 1;288(Pt 2):345–349. doi: 10.1042/bj2880345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Greenberg M. E., Ziff E. B. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene. Nature. 1984 Oct 4;311(5985):433–438. doi: 10.1038/311433a0. [DOI] [PubMed] [Google Scholar]
  11. Hellénius M. L., de Faire U., Berglund B., Hamsten A., Krakau I. Diet and exercise are equally effective in reducing risk for cardiovascular disease. Results of a randomized controlled study in men with slightly to moderately raised cardiovascular risk factors. Atherosclerosis. 1993 Oct;103(1):81–91. doi: 10.1016/0021-9150(93)90042-s. [DOI] [PubMed] [Google Scholar]
  12. Hiatt W. R., Regensteiner J. G., Hargarten M. E., Wolfel E. E., Brass E. P. Benefit of exercise conditioning for patients with peripheral arterial disease. Circulation. 1990 Feb;81(2):602–609. doi: 10.1161/01.cir.81.2.602. [DOI] [PubMed] [Google Scholar]
  13. Iba T., Sumpio B. E. Morphological response of human endothelial cells subjected to cyclic strain in vitro. Microvasc Res. 1991 Nov;42(3):245–254. doi: 10.1016/0026-2862(91)90059-k. [DOI] [PubMed] [Google Scholar]
  14. Iba T., Sumpio B. E. Tissue plasminogen activator expression in endothelial cells exposed to cyclic strain in vitro. Cell Transplant. 1992;1(1):43–50. doi: 10.1177/096368979200100108. [DOI] [PubMed] [Google Scholar]
  15. Kubes P., Suzuki M., Granger D. N. Nitric oxide: an endogenous modulator of leukocyte adhesion. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4651–4655. doi: 10.1073/pnas.88.11.4651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Leaf C. D., Wishnok J. S., Hurley J. P., Rosenblad W. D., Fox J. G., Tannenbaum S. R. Nitrate biosynthesis in rats, ferrets and humans. Precursor studies with L-arginine. Carcinogenesis. 1990 May;11(5):855–858. doi: 10.1093/carcin/11.5.855. [DOI] [PubMed] [Google Scholar]
  17. Leone A. M., Palmer R. M., Knowles R. G., Francis P. L., Ashton D. S., Moncada S. Constitutive and inducible nitric oxide synthases incorporate molecular oxygen into both nitric oxide and citrulline. J Biol Chem. 1991 Dec 15;266(35):23790–23795. [PubMed] [Google Scholar]
  18. Letsou G. V., Rosales O., Maitz S., Vogt A., Sumpio B. E. Stimulation of adenylate cyclase activity in cultured endothelial cells subjected to cyclic stretch. J Cardiovasc Surg (Torino) 1990 Sep-Oct;31(5):634–639. [PubMed] [Google Scholar]
  19. Marsden P. A., Heng H. H., Scherer S. W., Stewart R. J., Hall A. V., Shi X. M., Tsui L. C., Schappert K. T. Structure and chromosomal localization of the human constitutive endothelial nitric oxide synthase gene. J Biol Chem. 1993 Aug 15;268(23):17478–17488. [PubMed] [Google Scholar]
  20. Marsden P. A., Schappert K. T., Chen H. S., Flowers M., Sundell C. L., Wilcox J. N., Lamas S., Michel T. Molecular cloning and characterization of human endothelial nitric oxide synthase. FEBS Lett. 1992 Aug 3;307(3):287–293. doi: 10.1016/0014-5793(92)80697-f. [DOI] [PubMed] [Google Scholar]
  21. Miller V. M., Vanhoutte P. M. Endothelium-dependent contractions to arachidonic acid are mediated by products of cyclooxygenase. Am J Physiol. 1985 Apr;248(4 Pt 2):H432–H437. doi: 10.1152/ajpheart.1985.248.4.H432. [DOI] [PubMed] [Google Scholar]
  22. Nakamura M., Itoh H., Ikeda C., Yanagisawa E., Miyazawa Y., Hatogai F., Iwadare M. The efficacy of aerobic exercise therapy on hypertensive patients with mild cardiac complications. Ann Acad Med Singapore. 1992 Jan;21(1):38–41. [PubMed] [Google Scholar]
  23. Nishida K., Harrison D. G., Navas J. P., Fisher A. A., Dockery S. P., Uematsu M., Nerem R. M., Alexander R. W., Murphy T. J. Molecular cloning and characterization of the constitutive bovine aortic endothelial cell nitric oxide synthase. J Clin Invest. 1992 Nov;90(5):2092–2096. doi: 10.1172/JCI116092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Paffenbarger R. S., Jr, Wing A. L., Hyde R. T., Jung D. L. Physical activity and incidence of hypertension in college alumni. Am J Epidemiol. 1983 Mar;117(3):245–257. doi: 10.1093/oxfordjournals.aje.a113537. [DOI] [PubMed] [Google Scholar]
  25. Palmer R. M., Moncada S. A novel citrulline-forming enzyme implicated in the formation of nitric oxide by vascular endothelial cells. Biochem Biophys Res Commun. 1989 Jan 16;158(1):348–352. doi: 10.1016/s0006-291x(89)80219-0. [DOI] [PubMed] [Google Scholar]
  26. Palmer R. M., Rees D. D., Ashton D. S., Moncada S. L-arginine is the physiological precursor for the formation of nitric oxide in endothelium-dependent relaxation. Biochem Biophys Res Commun. 1988 Jun 30;153(3):1251–1256. doi: 10.1016/s0006-291x(88)81362-7. [DOI] [PubMed] [Google Scholar]
  27. Patel D. J., Greenfield J. C., Jr, Austen W. G., Morrow A. G., Fry D. L. Pressure-flow relationships in the ascending aorta and femoral artery of man. J Appl Physiol. 1965 May;20(3):459–463. doi: 10.1152/jappl.1965.20.3.459. [DOI] [PubMed] [Google Scholar]
  28. Poehlman E. T., Gardner A. W., Ades P. A., Katzman-Rooks S. M., Montgomery S. M., Atlas O. K., Ballor D. L., Tyzbir R. S. Resting energy metabolism and cardiovascular disease risk in resistance-trained and aerobically trained males. Metabolism. 1992 Dec;41(12):1351–1360. doi: 10.1016/0026-0495(92)90107-l. [DOI] [PubMed] [Google Scholar]
  29. Pollock J. S., Nakane M., Buttery L. D., Martinez A., Springall D., Polak J. M., Förstermann U., Murad F. Characterization and localization of endothelial nitric oxide synthase using specific monoclonal antibodies. Am J Physiol. 1993 Nov;265(5 Pt 1):C1379–C1387. doi: 10.1152/ajpcell.1993.265.5.C1379. [DOI] [PubMed] [Google Scholar]
  30. Posner J. D., Gorman K. M., Gitlin L. N., Sands L. P., Kleban M., Windsor L., Shaw C. Effects of exercise training in the elderly on the occurrence and time to onset of cardiovascular diagnoses. J Am Geriatr Soc. 1990 Mar;38(3):205–210. doi: 10.1111/j.1532-5415.1990.tb03492.x. [DOI] [PubMed] [Google Scholar]
  31. Radomski M. W., Palmer R. M., Moncada S. Comparative pharmacology of endothelium-derived relaxing factor, nitric oxide and prostacyclin in platelets. Br J Pharmacol. 1987 Sep;92(1):181–187. doi: 10.1111/j.1476-5381.1987.tb11310.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Resnick N., Collins T., Atkinson W., Bonthron D. T., Dewey C. F., Jr, Gimbrone M. A., Jr Platelet-derived growth factor B chain promoter contains a cis-acting fluid shear-stress-responsive element. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4591–4595. doi: 10.1073/pnas.90.10.4591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Robinson L. J., Weremowicz S., Morton C. C., Michel T. Isolation and chromosomal localization of the human endothelial nitric oxide synthase (NOS3) gene. Genomics. 1994 Jan 15;19(2):350–357. doi: 10.1006/geno.1994.1068. [DOI] [PubMed] [Google Scholar]
  34. Rosales O. R., Sumpio B. E. Changes in cyclic strain increase inositol trisphosphate and diacylglycerol in endothelial cells. Am J Physiol. 1992 Apr;262(4 Pt 1):C956–C962. doi: 10.1152/ajpcell.1992.262.4.C956. [DOI] [PubMed] [Google Scholar]
  35. Rosales O. R., Sumpio B. E. Protein kinase C is a mediator of the adaptation of vascular endothelial cells to cyclic strain in vitro. Surgery. 1992 Aug;112(2):459–466. [PubMed] [Google Scholar]
  36. Schmidt H. H., Nau H., Wittfoht W., Gerlach J., Prescher K. E., Klein M. M., Niroomand F., Böhme E. Arginine is a physiological precursor of endothelium-derived nitric oxide. Eur J Pharmacol. 1988 Sep 13;154(2):213–216. doi: 10.1016/0014-2999(88)90101-x. [DOI] [PubMed] [Google Scholar]
  37. Sessa W. C., Harrison J. K., Barber C. M., Zeng D., Durieux M. E., D'Angelo D. D., Lynch K. R., Peach M. J. Molecular cloning and expression of a cDNA encoding endothelial cell nitric oxide synthase. J Biol Chem. 1992 Aug 5;267(22):15274–15276. [PubMed] [Google Scholar]
  38. Sessa W. C., Pritchard K., Seyedi N., Wang J., Hintze T. H. Chronic exercise in dogs increases coronary vascular nitric oxide production and endothelial cell nitric oxide synthase gene expression. Circ Res. 1994 Feb;74(2):349–353. doi: 10.1161/01.res.74.2.349. [DOI] [PubMed] [Google Scholar]
  39. Steinman D. A., Ethier C. R. The effect of wall distensibility on flow in a two-dimensional end-to-side anastomosis. J Biomech Eng. 1994 Aug;116(3):294–301. doi: 10.1115/1.2895733. [DOI] [PubMed] [Google Scholar]
  40. Sumpio B. E., Banes A. J. Prostacyclin synthetic activity in cultured aortic endothelial cells undergoing cyclic mechanical deformation. Surgery. 1988 Aug;104(2):383–389. [PubMed] [Google Scholar]
  41. Sumpio B. E., Widmann M. D. Enhanced production of endothelium-derived contracting factor by endothelial cells subjected to pulsatile stretch. Surgery. 1990 Aug;108(2):277–282. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES