Dobesilate enhances endothelial nitric oxide synthase‐activity in macro‐ and microvascular endothelial cells

Abstract

1 Dobesilate is used for normalizing vascular dysfunction in a number of diseases. In search for an eect on endothelial NO production, macrovascular endothelial cells from rat aorta, microvascular endothelial cells from rat exocrine pancreatic tissue, and capillary endothelial cells from rat islets, were cultured in the presence or absence of Mg-Dobesilate. The activity of constitutive nitric oxide synthase (ecNOS) in resident cells as well as of inducible nitric oxide synthase (iNOS) in cytokine-activated cells was measured indirectly by recording the citrulline concentrations in culture supernatants. 2 In each of the dierent endothelial cells Mg-Dobesilate incubation (0.25 ± 1 mM) for 24 h led to a signi®cant and concentration-dependent increase in ecNOS-activities. With cytokine-activated endothelial cell cultures only moderate eects were seen with little or no concentration-dependency. Addition of the NOS-inhibitor N G -monomethyl-L-arginine led to a signi®cant suppression of citrulline formation in all cultures as an evidence for the enzyme speci®city of these eects. 3 iNOS-and ecNOS-speci®c reverse transcription and semi-quantitative polymerase chain reaction (RT ± PCR) with RNA from resident or cytokine-activated endothelial cells gave no evidence for an increase in NOS-speci®c mRNA after Mg-Dobesilate-treatment. Furthermore, Dobesilate-mediated enhancement of NO synthesis in resting endothelial cells was not due to iNOS induction in these cells, as no iNOS-speci®c signal was found by RT ± PCR.

p21 Cip1 Levels Differentially Regulate Turnover of Mature Endothelial Cells, Endothelial Progenitor Cells, and In Vivo Neovascularization

“…19,20 After 10 days in culture over 2 to 3 passages, 65Ϯ3% or 56Ϯ8% of the cells were positive for the EC-specific surface markers Flk-1/KDR/VEGF-R2 receptor tyrosine kinase or CD146 (S-Endo-1; MUC-18), respectively ( Figure 1A). Western blot analysis of whole cell lysates confirmed the absence of p21 protein from ECs derived from homozygous p21 Ϫ/Ϫ mice, whereas p21 protein levels were significantly reduced in cells from heterozygous p21 ϩ/Ϫ mice ( Figure 1B).…”

“…19,20 In brief, aortas from female or male mice were cleaned, longitudinally opened, and placed with the intimal side downward onto a collagen matrix in a medium supplemented with 10% FCS and 100 g/mL endothelial cell growth supplement (Calbiochem) for 6 days. After digestion of the matrix, the remaining cells were pelleted, seeded on fibronectin-coated 24-well plates, and grown to confluence in endothelial basal medium (EBM; Cell Systems).…”

Aging Enhances the Sensitivity of Endothelial Cells Toward Apoptotic Stimuli

“…22 Six-month-old, male wild-type and eNOS knockout mice (Jackson Laboratories) were killed and the aorta was removed. The vessel was cleaned of periadventitial fat and connective tissue, opened longitudinally, and placed with the aortic intimal side down into collagen-matrix-coated 24-well plates (collagen-matrix components: 3 mg/mL collagen, 5ϫDMEM, and 100ϫ penicillin/streptomycin).…”

Modern approach to early diagnosis and pathogenetic treatment of diabetic retinopathy

“…Отмечено, что у крыс с индуцированным стрептозотоцином СД (СТЗ-СД) длительное лечение задерживало прогрессирование ДР [9]. Важным механизмом действия КД является оптимизация эндотелиально-зависимой вазодила-тации (оксидазотзависимая) [10,11]. Изучены анти-оксидантные и противовоспалительные свойства КД.…”