This Article Abstract Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hishikawa, K. Articles by Saruta, T. Search for Related Content PubMed PubMed Citation Articles by Hishikawa, K. Articles by Saruta, T. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Hazardous Substances DB NITRIC OXIDE

(Hypertension. 1995;25:449-452.)
© 1995 American Heart Association, Inc.

Articles

Pressure Enhances Endothelin-1 Release From Cultured Human Endothelial Cells

Keiich Hishikawa; Toshio Nakaki; Takeshi Marumo; Hiromichi Suzuki; Ryuichi Kato; Takao Saruta

From the Departments of Internal Medicine and Pharmacology (T.N., R.K.), Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.

Correspondence to Toshio Nakaki, MD, PhD, Department of Pharmacology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160 Japan.

	Abstract

Top Abstract Introduction Methods Results Discussion References

 
Abstract The effect of pure pressure without shear stress or stretch on the release of endothelin-1 was investigated. Elevation of pressure significantly enhanced endothelin-1 release from cultured human umbilical vein endothelial cells. A calcium channel blocker, nifedipine, and a putative stretch-activated channel blocker, gadolinium, did not affect the pressure-induced endothelin-1 increase. On the other hand, a phospholipase C inhibitor, 2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate, and protein kinase C inhibitors, 1-5-(isoquinolinylsulfonyl)-2-methylpiperazine and chelerythrine, significantly inhibited the pressure-induced endothelin-1 increase. Moreover, pure pressure reduced basal nitric oxide release, while pretreatment with a nitric oxide synthase inhibitor, N^G-monomethyl-L-arginine, had no effect on the pressure-induced endothelin-1 increase. In conclusion, our results show for the first time that pressure enhances endothelin-1 release partially through activation of phospholipase C and protein kinase C in human endothelial cells.

Key Words: endothelin-1 • mechanoreception • Ca²⁺ channel • stretch • pressure • nitric oxide

	Introduction

Top Abstract Introduction Methods Results Discussion References

 
Endothelin-1 (ET-1), a 21–amino acid peptide produced primarily in endothelial cells, is the most powerful endogenous vasoconstrictor agent,¹ but the role of ET-1 in hypertension is still unclear. Plasma ET-1 concentrations in experimental and human hypertension are similar to or slightly higher than those in normotensive control subjects.^{2 3 4 5} Therefore, it is important to clarify the regulatory mechanism of ET-1 production by endothelial cells. It has been become clear recently that hemodynamic forces play an important role in the structure and function of vascular endothelial cells. The physical forces exerted on the blood vessel wall by the passage of intraluminal blood are pressure and shear stress. Pressure is exerted radially at right angles to the axis of flow, leads to tangential strain on the wall, and secondarily causes stretch. Shear stress, on the other hand, is exerted in the same axis as flow. The effect of shear stress on the release of various vasoactive substances^{6 7 8} has been investigated extensively, while the effect of pressure itself is not well understood. We recently devised a new method of studying pressure effects and reported that transmural pressure inhibits nitric oxide (NO) release from human endothelial cells.⁹ It was also found that pressure increases the production of 1,4,5-inositol trisphosphate in rat cultured vascular smooth muscle cells.^{10 11} This study was designed to investigate whether and how pressure modulates ET-1 release from human endothelial cells.

	Methods

Top Abstract Introduction Methods Results Discussion References

 
Materials
Nifedipine, 2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate (NCDC), and N^G-monomethyl-L-arginine (NMMA) were purchased from Sigma Chemical Co. Gadolinium chloride hexahydrate, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7), and N-(2-guanidinoethyl)-5-isoquinolinesulfonamide hydrochloride (HA1004) were purchased from Seikagaku Kogyo Co. Chelerythrine was purchased from LC Laboratories. Fetal calf serum (FCS) was purchased from Mitsubishi-Kasei. Endothelial cell growth supplement was purchased from Sigma Chemical Co. Collagen-coated 24-well plates were purchased from Corning and 175-cm² film panel flasks (EZIN Flasks) were purchased from Nunc Inc.

Cells
Human umbilical vein endothelial cells (HUVEC) were prepared from human umbilical cord veins according to the method described by Jaffe et al, with some minor modifications.⁹ The study protocol was previously approved by the ethics committee of the Keio University Hospital. HUVEC were cultured on collagen-coated 24-well plates in M199 medium supplemented with 10% FCS, 30 µg/mL endothelial cell growth supplement, and 6 U/mL heparin at 37°C under 5% CO₂ in air. Cells harvested between passages 3 and 5 were used for the experiments. At the end of each experiment, the 24-well plates were washed with Hanks' solution, and HUVEC were detached with 1% trypsin-EDTA. An aliquot of the suspension was counted to obtain the cell number with a Hemacytometer (Kayagaki Co). The result presented is an average of four fields. Cells were seeded with 2x10⁴ cells per well, and the number of cells did not change over the course of experiments.

Pressure-Loading Apparatus
A pressure-loading apparatus was set up as previously reported^{9 10 11} with some modification. Collagen-coated 24-well plates were placed in a film panel flask by peeling off the upper plastic film, and the flask was then tightly resealed. The flasks were tightly clamped between two iron plates, and the top of each flask was sealed with a rubber cap. The rubber cap was pierced by a needle connected to tubing attached to a three-way rotary valve, a sphygmomanometer, and a pressure valve. Compressed helium gas was pumped in to raise the internal pressure. Cell number was calculated at the end of each experiment, but there were no significant changes among experiments. Cell viability determined by trypan blue exclusion test at the end of the studies exceeded 90% in each experiment. Lactate dehydrogenase enzymatic activity was examined in some experiments, but no significant changes were found.

Assay of ET-1
Confluent cells were incubated for 2 to 8 hours in 1 mL serum-free medium containing various kinds of drugs or vehicle under various pressures. After incubation, the medium was collected in tubes and stored at -80°C until assay. The concentration of ET-1 was measured with a sandwich-enzyme immunoassay kit (IBL 17121, Immuno Bio Laboratory),¹² which detects only intact, active protein of ET-1.

Assay of NO
HUVEC were incubated for 8 hours in serum-free medium at atmospheric pressure and at 80 mm Hg. After incubation, the medium was collected and stored at -20°C until assay. NO production was evaluated by measurement of the stable end products of NO, NO₂^-, and NO₃^- with the Griess reaction, as described previously.¹³

Statistical Analysis
Results are expressed as mean±SEM. The statistical significance of the results was evaluated with ANOVA, and probability values were determined by Student's t test.

	Results

Top Abstract Introduction Methods Results Discussion References

 
Effect of Pressure on ET-1 Production From HUVEC
When confluent HUVEC were incubated with serum-free medium for 8 hours at atmospheric pressure and at 80 mm Hg, ET-1 production increased linearly with time (Fig 1). The pressure-induced increase in ET-1 release from HUVEC was enhanced by elevation of the pressure from 40 to 160 mm Hg in a pressure-dependent manner (Fig 2).

View larger version (14K): [in this window] [in a new window]   Figure 1. Line graph shows endothelin-1 (ET-1) concentration from cultured human umbilical vein endothelial cells at atmospheric pressure () and at 80 mm Hg () as a function of time. Each point represents the value obtained from a separate well. Values are mean±SEM (n=8). *P<.05 vs the value of 0 mm Hg at each time.

View larger version (36K): [in this window] [in a new window]   Figure 2. Bar graph shows dose-response curve for the pressures at which endothelin-1 (ET-1) release was observed. Each pressure-induced ET-1 increase was defined by subtracting values obtained at 0 mm Hg from those with applied pressure. Values are mean±SEM (n=8). *P<.05 vs the value of 40 mm Hg at each time.

Role of Voltage-Dependent Ca²⁺ Channel and Stretch-Activated Channel in Pressure-Induced ET-1 Increase
HUVEC were pretreated with a voltage-dependent Ca²⁺ channel blocker, nifedipine, and an inhibitor of stretch-activated channels, gadolinium, and then pressurized for 8 hours at 80 mm Hg. The level of ET-1 in the medium was determined. Each pressure-induced ET-1 increase was defined by subtraction of values obtained at 0 mm Hg from those with applied pressure. Nifedipine showed no effect on the pressure-induced ET-1 increase (Fig 3). Gadolinium, in the 1 to 10 µmol/L range, reportedly produces dose-dependent inhibition of stretch-activated current^{14 15} and stretch-induced arrythmias.¹⁶ We initially applied a 10-µmol concentration, but this caused cell death in just over half the HUVEC at 80 mm Hg. Therefore, we used 1 or 3 µmol/L. Neither of these concentrations affected the pressure-induced ET-1 increase (Fig 3).

View larger version (15K): [in this window] [in a new window]   Figure 3. Bar graph shows the effects of nifedipine and gadolinium on the pressure-induced endothelin-1 (ET-1) increase. Each pressure-induced ET-1 increase was defined by subtracting values obtained at 0 mm Hg from those with applied pressure. Values are mean±SEM (n=8). Vehicle indicates incubated with medium only; Nif, incubated with nifedipine (5 µmol/L) at 80 mm Hg; Gd 1, incubated with 1 µmol/L gadolinium at 80 mm Hg; and Gd 3, incubated with 3 µmol/L gadolinium at 80 mm Hg.

Role of Phospholipase C and Protein Kinase C in Pressure-Induced ET-1 Increase
To clarify the contribution of phospholipase C in the pressure-induced ET-1 increase, HUVEC were pretreated with NCDC and then pressurized for 8 hours at 80 mm Hg. Pretreatment with NCDC significantly inhibited the pressure-induced ET-1 increase at 80 mm Hg (Fig 4). Furthermore, to investigate the contribution of protein kinase C in this pressure-induced ET-1 increase, HUVEC were pretreated with protein kinase C inhibitor. Pretreatment with H-7 significantly inhibited the pressure-induced ET-1 increase at 80 mm Hg compared with an equimolar amount of the nonspecific protein kinase inhibitor HA1004. Chelerythrine chloride (0.6 µmol/L), a selective protein kinase C inhibitor, also significantly inhibited the pressure-induced ET-1 increase.

View larger version (22K): [in this window] [in a new window]   Figure 4. Bar graph shows the effects of phospholipase C and protein kinase C inhibitors on pressure-induced endothelin-1 (ET-1) increase. Each pressure-induced ET-1 increase was defined by subtracting values obtained at 0 mm Hg from those with applied pressure. Vehicle indicates 0.1% dimethyl sulfoxide (DMSO); NCDC, incubated with 2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate (200 µmol/L) under 80 mm Hg; HA, incubated with HA1004 (30 µmol/L) under 80 mm Hg; H7, incubated with H-7 (30 µmol/L) under 80 mm Hg; and chele, incubated with chelerythrine (0.6 µmol/L) at 80 mm Hg.

Role of NO in Pressure-Induced ET-1 Increase
To investigate the contribution of NO in pressure-induced NO release, NO and ET-1 release were measured simultaneously both under atmospheric conditions and after 80 mm Hg for 8 hours. Basal NO release was significantly inhibited by pressurization at 80 mm Hg (Fig 5A). Although pretreatment with NMMA virtually abolished NO release under atmospheric conditions and at 80 mm Hg, it had no effect on ET-1 release under either condition (Fig 5B).

View larger version (19K): [in this window] [in a new window]   Figure 5. Bar graphs show the effects of pressure and 500 µmol/L of NG-L-monomethyl-arginine (NMMA) on (A) nitric oxide (NO) and (B) endothelin-1 (ET-1) release. Human umbilical vein endothelial cells were incubated both under atmospheric conditions and at 80 mm Hg, with and without NMMA, for 8 hours. *P<.05 vs atmospheric condition without NMMA. **P<.05 vs atmospheric condition without NMMA. ***P<.05 vs atmospheric condition with NMMA.

	Discussion

Top Abstract Introduction Methods Results Discussion References

 
The major finding of the present study is that pressure elevation enhances ET-1 release from HUVEC. It is possible that the pressure-induced ET-1 increases are attributable simply to an increase in cell number. In fact, Tokunaga and Watanabe¹⁷ and Sumpio et al¹⁸ reported that 48 hours of pressurization increased endothelial cell numbers. We also recently reported that 24 to 48 hours of pressurization increased cell numbers in cultures of rat vascular smooth muscle cells.¹¹ There were, however, no significant differences in cell number among the experiments conducted in the present study. Eight hours may be too short a period to directly cause cell proliferation in response to pressure. These results show that the pressure-induced ET-1 increase was not caused by an increase in cell number. Furthermore, our results suggest that pressure causes proliferation of endothelial cells not only directly but also by a secondary effect, a growth factor¹⁹ mediated by the pressure-induced ET-1 increase.

Although confluent HUVEC attached to a solid plate can be subjected to pure pressure with minimal participation of pressure-induced stretch, we cannot completely exclude the possibility that pressure-induced stretch occurred. To clarify the contribution of stretch in pressure-induced ET-1 release, HUVEC were pretreated with gadolinium, a putative blocker of stretch-activated channels that has a calcium channel blocking effect. Gadolinium (1 to 3 µmol/L) had no effect at either dose used. Agonist-mediated stimulation of ET-1 release has been shown to be mediated by increased intracellular calcium and activation of protein kinase C pathways.^{20 21} To test the possibility that pressure-induced ET-1 release is mediated by Ca²⁺ channel modulation, HUVEC were also pretreated with a calcium channel blocker before being pressurized. We were unable to demonstrate any inhibition of pressure-induced ET-1 release with nifedipine. These results clearly show that the pressure-induced ET-1 increase is mediated by neither stretch-activated channels nor voltage-dependent Ca²⁺ channels and that the pressure produced with our method is different from stretch.

We recently demonstrated that pressure causes phosphoinositide turnover in vascular smooth muscle cells.^{10 11} In addition, shear stress–induced ET-1 release has been reported to be regulated by protein kinase C.⁸ We have hypothesized that pressure-induced ET-1 release is regulated by a pressure-induced phosphoinositide turnover and activation of protein kinase C. NCDC, a serine esterase inhibitor, has a well-defined specificity for the active site of phospholipase C and inhibits phosphoinositide-specific phospholipase C.²² NCDC significantly inhibited pressure-induced ET-1 release. The K_i of H-7 for protein kinase G, A, and C are 5.8, 3.0, and 6.0 µmol/L, respectively. The K_i of HA1004 for protein kinase G, A, and C are 1.3, 2.3, and 40 µmol/L, respectively. The pressure-induced ET-1 increase was significantly inhibited by 30 µmol/L of H-7 but not by HA1004. Chelerythrine chloride, a selective protein kinase C inhibitor, significantly inhibited pressure-induced ET-1 release at 80 mm Hg but could not completely inhibit pressure-induced ET-1 release. These results show that the pressure-induced ET-1 increase may be mediated partially by the activation of phospholipase C and protein kinase C. The possibility of mediation by another pathway, however, such as activation of tyrosine kinase¹¹ cannot be ruled out.

NO released during stimulation with thrombin has been reported not only to antagonize the effect of ET-1 but also to inhibit the agonist-stimulated production of ET-1 in endothelial cells.²³ To investigate the role of NO in pressure-induced ET-1 release, HUVEC were pressurized at 80 mm Hg for 8 hours, and NO and ET-1 release was measured simultaneously. Basal NO production was significantly inhibited by pressure, as in the case of histamine-stimulated NO release.⁹ Pretreatment with NMMA, an NO synthase inhibitor, virtually abolished NO release both under atmospheric conditions and at 80 mm Hg but had no effect on ET-1 release under either condition. These results indicate that the pressure-induced ET-1 increase is not secondarily enhanced by reduced production of basal NO. The basal level of NO may be too low to inhibit ET-1 release, possibly accounting for the fact that NO inhibition has no effect on ET-1 release. Recently, Marsden et al²⁴ reported that an AP-2–like element²⁵ exists in human endothelial NO synthase gene. Taken together, these results suggest that a pressure-induced ET-1 increase may be attributable partially to activation of protein kinase C and that basal NO release declines after inactivation of NO synthase owing to protein kinase C activation.

In the hypertensive state, the endothelial layer is exposed to mechanical stresses such as shear stress, stretch, and pressure. Pressure has been reported to cause cell proliferation^{17 18} and to modulate prostacyclin synthesis¹⁷ in endothelial cells. The pressure-mediated pathway, however, has been poorly understood. Although the contribution of a protease,²⁶ which inactivates ET-1, to the pressure-induced ET-1 increase cannot be ruled out, our results show for the first time that pressure enhances ET-1 release by partially activating phospholipase C and protein kinase C rather than by activating calcium- and stretch-activated channels. These results support the concept that not only shear stress and stretch but also pressure, as a mechanical stress, play an important role in regulating endothelial function.

	Acknowledgments

 
This work was supported in part by the Mitsukoshi Prize of Medicine 1992 and by a grant-in-aid for general scientific research from the Ministry of Education, Science and Culture of Japan.

Received May 24, 1994; first decision July 22, 1994; accepted November 29, 1994.

	References

Top Abstract Introduction Methods Results Discussion References

 

1. Yanagisawa M, Kurihara H, Kimura S, Tomobe Y, Kobayashi M, Mitsui Y, Yazaki Y, Goto K, Masaki T. A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature. 1988;332:411-415. [Medline] [Order article via Infotrieve]
2. Nguyen PV, Parent A, Deng LY, Flückiger JP, Thibault G, Schiffrin EL. Endothelin vascular receptors and responses in DOCA-salt hypertensive rats. Hypertension. 1992;19(suppl II):II-98-II-104.
3. Suzuki N, Miyauchi T, Tomobe Y, Matsumoto H, Goto K, Masaki T, Fujimo M. Plasma concentrations of endothelin-1 in spontaneously hypertensive rats and DOCA-salt hypertensive rats. Biochem Biophys Res Commun. 1990;167:941-947. [Medline] [Order article via Infotrieve]
4. Schiffrin EL, Thibault G. Plasma endothelin in human essential hypertension. Am J Hypertens. 1991;4:303-308. [Medline] [Order article via Infotrieve]
5. Kohno M, Murakawa K, Horio T, Yokokawa K, Tasunari K, Fukui T, Takeda T. Plasma immunoreactive endothelin-1 in experimental malignant hypertension. Hypertension. 1991;18:93-100. [Abstract/Free Full Text]
6. Frangos JA, Eskin SG, McIntire LV, Ives CL. Flow effects on prostacyclin production by cultured human endothelial cells. Science. 1985;227:1477-1479. [Abstract/Free Full Text]
7. Buga GM, Gold ME, Fukuto JM, Ignarro LJ. Shear stress-induced release of nitric oxide from endothelial cells grown on beads. Hypertension. 1991;17:187-193. [Abstract/Free Full Text]
8. Kuchan MJ, Frangos JA. Shear stress regulates endothelin-1 release via protein kinase C and cGMP in cultured endothelial cells. Am J Physiol. 1993;33:H150-H156.
9. Hishikawa K, Nakaki T, Suzuki H, Saruta T, Kato R. Transmural pressure inhibits nitric oxide release from human endothelial cells. Eur J Pharmacol. 1992;215:329-331. [Medline] [Order article via Infotrieve]
10. Hishikawa K, Nakaki T, Suzuki H, Kato R, Saruta T. Pressure-sensitive mechanoreception in endothelial cells and smooth muscle cells. Hypertension. 1992;20:424. Abstract.
11. Hishikawa K, Nakaki T, Marumo T, Hayashi M, Suzuki H, Kato R, Saruta T. Pressure promotes DNA synthesis in rat cultured smooth muscle cells. J Clin Invest. 1994;93:1975-1980.
12. Suzuki N, Matsumoto H, Kitada C, Masaki T, Fujino M. A sensitive sandwich-enzyme immunoassay for human endothelin. J Immunol Methods. 1989;118:245-250. [Medline] [Order article via Infotrieve]
13. Hishikawa K, Nakaki T, Tsuda M, Esumi H, Ohshima H, Suzuki H, Saruta T, Kato R. Effect of systemic L-arginine administration on hemodynamics and nitric oxide release in man. Jpn Heart J. 1992;33:41-48. [Medline] [Order article via Infotrieve]
14. Yang XC, Sachs F. Block of stretch-activated ion channels in xenopus oocytes by gadolinium and calcium ions. Science. 1989;243:1068-1071. [Abstract/Free Full Text]
15. Docherty RJ. Gadolinium selectively blocks a component of calcium current in rodent neuroblastoma X glioma hybrid (NG108-15) cells. J Physiol (Lond). 1988;398:33-47. [Abstract/Free Full Text]
16. Hansen DE, Borganelli M, Stacy GPJ, Taylor LK. Dose-dependent inhibition of stretch-induced arrhythmias by gadolinium in isolated canine ventricles. Circ Res. 1991;69:820-831. [Abstract/Free Full Text]
17. Tokunaga O, Watanabe T. Properties of endothelial cell and smooth muscle cell cultured in ambient pressure. In Vitro Cell Dev Biol. 1987;23:528-534. [Medline] [Order article via Infotrieve]
18. Sumpio BE, Widmann MD, Ricotta J, Awolesi MA, Watase M. Increased ambient pressure stimulates proliferation and morphologic changes in cultured endothelial cells. J Cell Physiol. 1994;158:133-139. [Medline] [Order article via Infotrieve]
19. Nakaki T, Nakayama M, Yamamoto S, Kato R. Endothelin-mediated stimulation of DNA synthesis in vascular smooth muscle cells. Biochem Biophys Res Commun. 1989;158:880-883. [Medline] [Order article via Infotrieve]
20. Yanagisawa M, Inoue A, Takuwa Y, Mitsui Y, Kobayashi M, Masaki T. The human preproendothelin-1 gene: possible regulation by endothelial phosphoinositide turnover signalling. J Cardiovasc Pharmacol. 1989;13:S13-S17.
21. Emori T, Hirata Y, Ohta K, Kanno K, Eguchi S, Imai T, Shirai M, Marumo F. Cellular mechanism of endothelin-1 release by angiotensin and vasopressin. Hypertension. 1991;18:165-170. [Abstract/Free Full Text]
22. Nakaki T, Roth BL, Chuang DM, Costa E. Phasic and tonic components in 5HT₂ receptor-mediated rat aorta contraction: participation of Ca²⁺ channels and phospholipase C. J Pharmacol Exp Ther. 1985;234:442-446. [Abstract/Free Full Text]
23. Boulanger C, Lüscher TF. Release of endothelin from the porcine aorta: inhibition by endothelium-derived nitric oxide. J Clin Invest. 1990;85:587-590.
24. Marsden PA, Heng HHQ, Scherer SW, Stewart RJ, Hall AV, Shi XM, Tsui LP, Schappert KT. Structure and chromosomal localization of the human constitutive endothelial nitric oxide synthase gene. J Biol Chem. 1993;23:17478-17488.
25. Imagawa M, Chiu R, Karin M. Transcription factor AP-2 mediates induction by two different signal-transduction pathways: protein kinase C and cAMP. Cell. 1987;51:251-260. [Medline] [Order article via Infotrieve]
26. Jackman HL, Morris PW, Rabito SF, Johansson GB, Skidgel RA, Erdros EG. Inactivation of endothelin-1 by an enzyme of the vascular endothelial cells. Hypertension. 1993;21:925-928.[Abstract/Free Full Text]

This article has been cited by other articles:

				J. L. Garvin and N. J. Hong Cellular Stretch Increases Superoxide Production in the Thick Ascending Limb Hypertension, February 1, 2008; 51(2): 488 - 493. [Abstract] [Full Text] [PDF]

				H. A. Lee, E. B. Baek, K. S. Park, H. J. Jung, J. I. Kim, S. J. Kim, and Y. E Earm Mechanosensitive nonselective cation channel facilitation by endothelin-1 is regulated by protein kinase C in arterial myocytes Cardiovasc Res, November 1, 2007; 76(2): 224 - 235. [Abstract] [Full Text] [PDF]

				K. P. Conrad Mechanisms of Renal Vasodilation and Hyperfiltration During Pregnancy Reproductive Sciences, October 1, 2004; 11(7): 438 - 448. [Abstract] [PDF]

				K. P. Conrad and J. Novak Emerging role of relaxin in renal and cardiovascular function Am J Physiol Regulatory Integrative Comp Physiol, August 1, 2004; 287(2): R250 - R261. [Abstract] [Full Text] [PDF]

				A. B. C. Coumans, Y. Garnier, S. Supcun, A. Jense, R. Berger, and T. H. M. Hasaart The Effects of Low-Dose Endotoxin on the Umbilicoplacental Circulation in Preterm Sheep Reproductive Sciences, July 1, 2004; 11(5): 289 - 293. [Abstract] [PDF]

				Y. Garnier, A. B. C. Coumans, A. Jensen, T. H. M. Hasaart, and R. Berger Infection-Related Perinatal Brain Injury: The Pathogenic Role of Impaired Fetal Cardiovascular Control Reproductive Sciences, December 1, 2003; 10(8): 450 - 459. [Abstract] [PDF]

				K. Minamoto, D. J. Pinsky, T. Fujita, and Y. Naka Timing of Nitric Oxide Donor Supplementation Determines Endothelin-1 Regulation and Quality of Lung Preservation for Transplantation Am. J. Respir. Cell Mol. Biol., January 1, 2002; 26(1): 14 - 21. [Abstract] [Full Text] [PDF]

				Y. Dumont, M. D'Amours, M. Lebel, and R. Lariviere Supplementation with a low dose of l-arginine reduces blood pressure and endothelin-1 production in hypertensive uraemic rats Nephrol. Dial. Transplant., April 1, 2001; 16(4): 746 - 754. [Abstract] [Full Text] [PDF]

				L.-m. Gan, L. Selin-Sjogren, R. Doroudi, and S. Jern Temporal regulation of endothelial ET-1 and eNOS expression in intact human conduit vessels exposed to different intraluminal pressure levels at physiological shear stress Cardiovasc Res, October 1, 2000; 48(1): 168 - 177. [Abstract] [Full Text] [PDF]

				Y. Kusaka, R. A. Kelly, G. H. Williams, and I. Kifor Coronary microvascular endothelial cells cosecrete angiotensin II and endothelin-1 via a regulated pathway Am J Physiol Heart Circ Physiol, September 1, 2000; 279(3): H1087 - H1096. [Abstract] [Full Text] [PDF]

				I. S. Wittstein, W. Qiu, R. C. Ziegelstein, Q. Hu, and D. A. Kass Opposite Effects of Pressurized Steady Versus Pulsatile Perfusion on Vascular Endothelial Cell Cytosolic pH : Role of Tyrosine Kinase and Mitogen-Activated Protein Kinase Signaling Circ. Res., June 23, 2000; 86(12): 1230 - 1236. [Abstract] [Full Text] [PDF]

				A. M. Malek, S. L. Alper, and S. Izumo Hemodynamic Shear Stress and Its Role in Atherosclerosis JAMA, December 1, 1999; 282(21): 2035 - 2042. [Abstract] [Full Text] [PDF]

				H. Xie, J. A. Bevan, and W. G. Mayhan Oxidized Low-Density Lipoprotein Enhances Myogenic Tone in the Rabbit Posterior Cerebral Artery Through the Release of Endothelin-1 • Editorial Comment Stroke, November 1, 1999; 30 (11): 2423 - 2430. [Abstract] [Full Text] [PDF]

				E. L. Schiffrin Role of Endothelin-1 in Hypertension Hypertension, October 1, 1999; 34(4): 876 - 881. [Abstract] [Full Text] [PDF]

				D. Henrion, M. Iglarz, and B. I. Levy Chronic Endothelin-1 Improves Nitric Oxide–Dependent Flow-Induced Dilation in Resistance Arteries From Normotensive and Hypertensive Rats Arterioscler. Thromb. Vasc. Biol., September 1, 1999; 19(9): 2148 - 2153. [Abstract] [Full Text] [PDF]

				E. Brochu, S. Lacasse-M., C. Moreau, M. Lebel, I. Kingma, J. H. Grose, and R. Lariviere Endothelin ETA receptor blockade prevents the progression of renal failure and hypertension in uraemic rats Nephrol. Dial. Transplant., August 1, 1999; 14(8): 1881 - 1888. [Abstract] [Full Text] [PDF]

				J. M. Tarbell, L. Demaio, and M. M. Zaw Effect of pressure on hydraulic conductivity of endothelial monolayers: role of endothelial cleft shear stress J Appl Physiol, July 1, 1999; 87(1): 261 - 268. [Abstract] [Full Text] [PDF]

				G. Marano, M. Grigioni, S. Palazzesi, and A. U Ferrari Endothelin and mechanical properties of the carotid artery in Wistar-Kyoto and spontaneously hypertensive rats Cardiovasc Res, March 1, 1999; 41(3): 701 - 707. [Abstract] [Full Text] [PDF]

				M. Iglarz, K. Matrougui, B. I. Levy, and D. Henrion Chronic blockade of endothelin ETA receptors improves flow dependent dilation in resistance arteries of hypertensive rats Cardiovasc Res, September 1, 1998; 39(3): 657 - 664. [Abstract] [Full Text] [PDF]

				R. A. Caldwell, H. F. Clemo, and C. M. Baumgarten Using gadolinium to identify stretch-activated channels: technical considerations Am J Physiol Cell Physiol, August 1, 1998; 275(2): C619 - C621. [Abstract] [Full Text] [PDF]

				A. Zanchi, N. Stergiopulos, H. R. Brunner, and D. Hayoz Differences in the Mechanical Properties of the Rat Carotid Artery In Vivo, In Situ, and In Vitro Hypertension, July 1, 1998; 32(1): 180 - 185. [Abstract] [Full Text] [PDF]

				T. Ziegler, K. Bouzourene, V. J. Harrison, H. R. Brunner, and D. Hayoz Influence of Oscillatory and Unidirectional Flow Environments on the Expression of Endothelin and Nitric Oxide Synthase in Cultured Endothelial Cells Arterioscler. Thromb. Vasc. Biol., May 1, 1998; 18(5): 686 - 692. [Abstract] [Full Text] [PDF]

				K. Hishikawa, B. S. Oemar, Z. Yang, and T. F. Luscher Pulsatile Stretch Stimulates Superoxide Production and Activates Nuclear Factor-{kappa}B in Human Coronary Smooth Muscle Circ. Res., November 19, 1997; 81(5): 797 - 803. [Abstract] [Full Text]

				K. Hishikawa and T. F. Luscher Pulsatile Stretch Stimulates Superoxide Production in Human Aortic Endothelial Cells Circulation, November 18, 1997; 96(10): 3610 - 3616. [Abstract] [Full Text]

				R. Largo, D. Gomez-Garre, X. H. Liu, J. Alonso, J. Blanco, J. J. Plaza, and J. Egido Endothelin-1 Upregulation in the Kidney of Uninephrectomized Spontaneously Hypertensive Rats and Its Modification by the Angiotensin-Converting Enzyme Inhibitor Quinapril Hypertension, May 1, 1997; 29(5): 1178 - 1185. [Abstract] [Full Text]

				T. Tonnessen, G. Christensen, E. Oie, E. Holt, H. Kjekshus, O. A. Smiseth, O. M. Sejersted, and H. Attramadal Increased cardiac expression of endothelin-1 mRNA in ischemic heart failure in rats Cardiovasc Res, March 1, 1997; 33(3): 601 - 610. [Abstract] [PDF]

				D. Hasdai, D. R. Holmes, K. N. Garratt, W. D. Edwards, and A. Lerman Mechanical Pressure and Stretch Release Endothelin-1 From Human Atherosclerotic Coronary Arteries In Vivo Circulation, January 21, 1997; 95(2): 357 - 362. [Abstract] [Full Text]

				D. Dunbar Ivy, J. P. Kinsella, and S. H. Abman Endothelin blockade augments pulmonary vasodilation in the ovine fetus J Appl Physiol, December 1, 1996; 81(6): 2481 - 2487. [Abstract] [Full Text] [PDF]

				P. R. Knoblich, R. H. Freeman, and D. Villarreal Pressure-Dependent Renin Release During Chronic Blockade of Nitric Oxide Synthase Hypertension, November 1, 1996; 28(5): 738 - 742. [Abstract] [Full Text]

				P. Sventek, J.-S. Li, K. Grove, C. F. Deschepper, and E. L. Schiffrin Vascular Structure and Expression of Endothelin-1 Gene in L-NAME–Treated Spontaneously Hypertensive Rats Hypertension, January 1, 1996; 27(1): 49 - 55. [Abstract] [Full Text]

				K. Hishikawa, B. S. Oemar, and T. Nakaki Static Pressure Regulates Connective Tissue Growth Factor Expression in Human Mesangial Cells J. Biol. Chem., May 11, 2001; 276(20): 16797 - 16803. [Abstract] [Full Text] [PDF]

This Article Abstract Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal