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Vascular Medicine
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Differential expression of nitric oxide by dermal microvascular endothelial cells from patients with scleroderma

Luz I Romero

Department of Dermatology Stanford University School of Medicine, Stanford, CA, USA

Dan-Ning Zhang

Department of Dermatology Stanford University School of Medicine, Stanford, CA, USA

John P Cooke

Department of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, CA, USA

Hoai-Ky V Ho

Department of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, CA, USA

Esperanza Avalos

Centro de Biología Experimental, Guadalupe, Zacatecas 98600, Mexico

Rafael Herrera

Centro de Biología Experimental, Guadalupe, Zacatecas 98600, Mexico

G Scott Herron

Department of Dermatology Stanford University School of Medicine, Stanford, CA, USA

Vascular abnormalities in scleroderma are fundamental to the pathogenesis of this disease. The objective of this study was to characterize dermal microvascular endothelial cells (DMEC) isolated from scleroderma patients with respect to growth and expression of the constitutive form of endothelial nitric oxide synthase (eNOS). DMEC from patients with both systemic sclerosis (SSc) and localized scleroderma (Loc Scl) contained small intact microvascular structures in contrast to single cell isolations obtained from control skin. Immunoaffinity selection on anti-PECAM-1 beads yielded pure populations of DMEC expressing normal markers. While the morphology and initial growth of SSc DMEC closely paralleled control cells, the growth of SSc DMEC decreased with time in culture (doubling time of 3 days vs. 5 days). Expression of ecNOS mRNA was reduced in both Loc Scl and SSc as shown by semi-quantitative RT-PCR (p, 0.001). Western blots showed variable but generally lower ecNOS protein levels and decreased levels of nitrogen oxides in media were found from both SSc and Loc Scl relative to control cells. The results indicate an intrinsic defect in the mechanism of nitric oxide production in DMEC isolated from scleroderma patients and suggest its possible involvement in the pathophysiology of scleroderma.

Key Words: constitutive endothelial-type nitric oxide synthase • human dermal microvascular endothelial cells • localized scleroderma • systemic scleroderma

Vascular Medicine, Vol. 5, No. 3, 147-158 (2000)
DOI: 10.1177/1358836X0000500304
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