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Vascular Bed Molecular Profiling by Differential Systemic Decellularization In Vivo.
Serra, Aida; Gallart-Palau, Xavier; Park, Jung Eun; Lim, Grace Gui Yin; Lim, Kah Leong; Ho, Hee Hwa; Tam, James P; Sze, Siu Kwan.
Afiliação
  • Serra A; From the School of Biological Sciences, Nanyang Technological University, Singapore (A.S., X.G.-P., J.E.P., J.P.T., S.K.S.).
  • Gallart-Palau X; From the School of Biological Sciences, Nanyang Technological University, Singapore (A.S., X.G.-P., J.E.P., J.P.T., S.K.S.).
  • Park JE; From the School of Biological Sciences, Nanyang Technological University, Singapore (A.S., X.G.-P., J.E.P., J.P.T., S.K.S.).
  • Lim GGY; Neurodegeneration Research Laboratory, National Neuroscience Institute, Singapore (G.G.Y.L., K.L.L.).
  • Lim KL; Neurodegeneration Research Laboratory, National Neuroscience Institute, Singapore (G.G.Y.L., K.L.L.).
  • Ho HH; Department of Physiology, National University of Singapore (K.L.L.).
  • Tam JP; Department of Cardiology, Tan Tock Seng Hospital, Singapore (H.H.H.).
  • Sze SK; From the School of Biological Sciences, Nanyang Technological University, Singapore (A.S., X.G.-P., J.E.P., J.P.T., S.K.S.).
Arterioscler Thromb Vasc Biol ; 38(10): 2396-2409, 2018 10.
Article em En | MEDLINE | ID: mdl-30354219
ABSTRACT
Objective- Vascular endothelial dysfunction is a key component of several major human diseases, but the molecular basis of this complex disorder has been difficult to determine in vivo. Previous attempts to identify key mediators of vascular endothelial dysfunction in experimental models have been limited by the lack of suitable methods for system-wide analyses of vascular bed biology. Here, we aimed to develop a novel method for investigating vascular endothelial dysfunction pathogenesis that enables system-wide analyses of molecular interactions between endothelial glycocalyx, endothelial cells, and smooth muscle cells in murine. Approach and Results- We developed a new technique using whole-body differential perfusion with increasing concentrations of detergent buffer to selectively solubilize distinct layers of vascular bed tissue in rodents. When combined with proteomics techniques, our novel approach of differential systemic decellularization in vivo enabled quantitative profiling of vascular beds throughout the body. Initial perfusion with phosphate buffer was used to obtain the endothelial glycocalyx, followed by subsequent extraction of endothelial cell components, and finally by smooth muscle cell constituents with increasing concentrations of detergent. Differential systemic decellularization in vivo has also been successfully applied to characterize molecular events in the vascular bed pathology of lipopolysaccharide-challenged mice. Conclusions- Together, these data indicate that differential systemic decellularization in vivo permits system-wide molecular characterization of vascular bed proteomes in rodent models and can be used to advance our current understanding of vascular endothelial dysfunction pathogenesis and progression in a wide range of disease settings.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Aorta Torácica / Perfusão / Endotoxemia / Proteoma / Proteômica / Ácido Desoxicólico / Detergentes Tipo de estudo: Prognostic_studies Limite: Animals Idioma: En Ano de publicação: 2018 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Aorta Torácica / Perfusão / Endotoxemia / Proteoma / Proteômica / Ácido Desoxicólico / Detergentes Tipo de estudo: Prognostic_studies Limite: Animals Idioma: En Ano de publicação: 2018 Tipo de documento: Article