Role of glutathione biosynthesis in endothelial dysfunction and fibrosis
| Autor: | Verónica Miguel, Susana Vallejo, Pablo Cannata, Eva Maria Blanco, Concepción Peiró, Elena Sandoval, Francisco Sánchez, Carlos F. Sánchez-Ferrer, Santiago Lamas, Cristina Espinosa-Diez |
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| Přispěvatelé: | UAM. Departamento de Anatomía Patológica, UAM. Departamento de Farmacología, Instituto de Investigación Sanitaria Fundación Jiménez Díaz (IIS-FJD), Instituto de Investigación Sanitaria Hospital Universitario de La Paz (IdiPAZ), Centro de Biología Molecular Severo Ochoa (CBM), Enogas, Fundación Ramón Areces, European Cooperation in Science and Technology, Comunidad de Madrid, Instituto de Salud Carlos III, European Commission, Fundación Renal Íñigo Álvarez de Toledo |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Male Clinical Biochemistry GAPDH glyceraldehyde 3-phosphate dehydrogenase NAC N-acetylcysteine Haploinsufficiency Kidney Biochemistry KHS Krebs-Henseleit solution chemistry.chemical_compound Mice EDRF Endothelium-derived relaxing factor Enos Fibrosis Kidney Fibrosis GCL glutamate cysteine ligase Endothelial dysfunction SDS sodium dodecyl sulphate lcsh:QH301-705.5 Cells Cultured chemistry.chemical_classification NA noradrenaline Mice Knockout lcsh:R5-920 biology Chemistry GCLM UUO Unilateral ureteral obstruction eNOS Endothelial nitric-oxide synthase ROS Biología y Biomedicina / Biología Farmacia Glutathione Mesenteric Arteries GCLC SNP sodium nitroprusside Female Kidney Diseases lcsh:Medicine (General) Research Paper Ach acetylcholine medicine.medical_specialty Nitric Oxide Synthase Type III BH4 Tetrahydrobiopterin Glutamate-Cysteine Ligase Biopterin ARE antioxidant response element Glutamate-cysteine ligase 03 medical and health sciences DMNQ 2 3-Dimethoxy-1 4-naphthoquinone Internal medicine medicine Animals Reactive oxygen species Organic Chemistry Endothelial Cells Fibroblasts biology.organism_classification medicine.disease MLEC mouse lung endothelial cells Disease Models Animal 030104 developmental biology Endocrinology lcsh:Biology (General) GS glutathione synthetase Reactive Oxygen Species |
| Zdroj: | Repositorio Institucional de la Consejería de Sanidad de la Comunidad de Madrid Consejería de Sanidad de la Comunidad de Madrid Biblos-e Archivo. Repositorio Institucional de la UAM instname Redox Biology Redox Biology, Vol 14, Iss, Pp 88-99 (2018) Digital.CSIC. Repositorio Institucional del CSIC |
| Popis: | Glutathione (GSH) biosynthesis is essential for cellular redox homeostasis and antioxidant defense. The rate-limiting step requires glutamate-cysteine ligase (GCL), which is composed of the catalytic (GCLc) and the modulatory (GCLm) subunits. To evaluate the contribution of GCLc to endothelial function we generated an endothelial-specific Gclc haplo-insufficient mouse model (Gclc e/+ mice). In murine lung endothelial cells (MLEC) derived from these mice we observed a 50% reduction in GCLc levels compared to lung fibroblasts from the same mice. MLEC obtained from haplo-insufficient mice showed significant reduction in GSH levels as well as increased basal and stimulated ROS levels, reduced phosphorylation of eNOS (Ser 1177) and increased eNOS S-glutathionylation, compared to MLEC from wild type (WT) mice. Studies in mesenteric arteries demonstrated impaired endothelium-dependent vasodilation in Gclc(e/+) male mice, which was corrected by pre-incubation with GSH-ethyl-ester and BH4. To study the contribution of endothelial GSH synthesis to renal fibrosis we employed the unilateral ureteral obstruction model in WT and Gclc(e/+) mice. We observed that obstructed kidneys from Gclc(e/+) mice exhibited increased deposition of fibrotic markers and reduced Nrf2 levels. We conclude that the preservation of endothelial GSH biosynthesis is not only critical for endothelial function but also in anti-fibrotic responses. Graphical abstract Specific endothelial reduction of GSH synthesis increases basal and stimulated ROS levels, reduces phosphorylation of eNOS (Ser 1177) and increases eNOS S-glutathionylation, impairing endothelium-dependent vasodilation and promoting renal fibrosis following UUO.fx1 Highlights • Endothelial haplo-insufficiency of GCLc increases basal and stimulated ROS levels. • Decreased endothelial GSH promotes eNOS S-glutathionylation. • Endothelial GSH reduction impairs endothelium-dependent vasodilation in male mice. • Reduced endothelial GSH levels promotes renal fibrosis. |
| Databáze: | OpenAIRE |
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