SOLANGE CARRASCO
Projetos de Pesquisa
Unidades Organizacionais
Departamento de Clínica Médica, Faculdade de Medicina
LIM/17 - Laboratório de Investigação em Reumatologia, Hospital das Clínicas, Faculdade de Medicina
LIM/17 - Laboratório de Investigação em Reumatologia, Hospital das Clínicas, Faculdade de Medicina
10 resultados
Resultados de Busca
Agora exibindo 1 - 10 de 10
conferenceObject Fibrogenesis failure of type V collagen observed in pulmonary and cutaneous fibroblast culture reinforce the pathogenic participation of this collagen in the pathway of systemic sclerosis(2012) TEODORO, W. R.; MORAIS, J.; MARTIN, P.; VELOSA, A. P. P.; CARRASCO, S.; SOUZA, R. B. C.; KATAYAMA, M. L.; GOLDEINSTEIN-SCHAINBERG, C.; PARRA, E. R.; CAPELOZZI, V. L.; YOSHINARI, N. H.Introduction: Unusual type V collagen (COLV) accumulation was demonstrated in systemic sclerosis (SSc) by our group. In this regard, this study analyzed tridimensional reconstruction (3D), biochemical and molecular profile of COLVα1 and COLVα2 chains in pulmonary and cutaneous fibroblasts culture from patients with SSc. Materials and Methods: Pulmonary and cutaneous fibroblasts for culture were obtained from 7 patients with SSc and from six controls respectively. COLV 3D reconstruction was performed by confocal microscopy. COLVα1 and COLVα2 gene expression was performed by RT-PCR and COLV protein expression by immunoblotting. Results: COL V 3D reconstruction showed distorted and strongly thickened fibers with irregular bundles resulting in a dense network in lung and skin fibroblast cultures from SSc patients compared to the thin fibers from fibroblast controls. Collagen quantification showed significant increased COLV fiber expression in SSc cutaneous and pulmonary fibroblasts (P<0.01) compared with the respective controls. In the same way, molecular evaluation demonstrated an increased significance (P=0.05) of COLVα1 and COLVα2 mRNA expression in cutaneous and pulmonary fibroblasts from SSc patients to that of control groups. The immunoblotting analysis demonstrated the increased weight of the molecular COLV chains. Conclusion: COLV overexpression and an unusual organization of these fibers including molecular and biochemical changes, suggest an interference process of the COLV fibrillogenesis in patients with SSc, reinforcing the participation of this collagen in SSc pathogenesis and open new therapeutic perspectives for these patients.conferenceObject INFLUENCE OF ALPHA 2 DO COLLAGEN V OVEREXPRESSION IN PHYSIOPHATOLOGY OF FIBROSIS SYSTEMIC SCLEROSIS PATIENTS(2014) MORAIS, J.; MARTIN, P.; VELOSA, A. P. P.; ANDRADE, P. C.; CRUZ, I. B.; MIRACCA, E. C.; FAC, F. A. C. Barrence; CARRASCO, S.; GOLDEINSTEIN-SCHAINBERG, C.; NAGAI, M. A.; PARRA, E. R.; CAPELOZZI, V. L.; TEODORO, W. R.conferenceObject Interstitial lung disease in systemic sclerosis is associated with autoimmunity to alpha 1(V) chain of type V collagen(2019) VELOSA, A. P. Pereira; BRITO, L.; QUEIROZ, Z. A.; CARRASCO, S.; MIRANDA, J. Tomaz de; GOLDENSTAIN-SCHAINBERG, C.; PARRA, E. Roger; CAPELOZZI, V. L.; TEODORO, W. RosoliaconferenceObject DYNAMIC COLLAGEN V REMODELING IS RELATED TO SKIN THICKENING IN SSc(2012) MARTIN, P.; TEODORO, W. R.; VELOSA, A. P.; CARRASCO, S.; MORAIS, J. de; CHRISTMANN, R. B.; PARRAS, E. R.; CAPELOZZI, V. L.; YOSHINARI, N. H.Background. Normal physiological properties of skin, one of the primary organs affected in SSc, depends on collagen Types I (COL I), III (COLIII) and V (COLV) assembly forming heterotypic fibres. COLV regulates fibril diameter and loss of this function could result in tissue fibrosis. In this way, our aim was to evaluate the histological and molecular profiles of COLI, COLIII and COLV in SSc skin and its correlation with skin thickening and disease activity. Methods. Skin biopsies of 18 patients (5 at early and 13 at late disease stage) and 10 healthy controls were studied. Assessment of skin thickening was performed using the modified Rodnan skin score (MRSS) and disease activity was calculated by Valentini Disease Activity Index. Quantification of COLI, COLIII and COLV was evaluated by histomorphometry in dermis and quantitative RT–PCR in dermal fibroblast culture. Results. A higher expression of abnormal COLV was observed in dermis of patients with early disease when compared with control group and late disease. The COLIII content was also higher in early SSc when compared with healthy controls and late SSc. On the other hand, the amount of COLI was higher in late disease when compared with control and early SSc. A positive correlation between COLV and MRSS (r = 0.42, P = 0.04) as well as disease activity (r = 0.45, P = 0.03) was observed, but there was no correlation between COLI and COLIII expression and these parameters. COLV α-1 and COLV α-2, as well as COLI α-1 and COLIII α-1 mRNA expression were higher in SSc when compared with control group. Conclusion. We found increased COLIII and COLV deposition in early SSc and increased COLI expression in late SSc indicating that collagen remodelling in SSc is a dynamic process. The fact that abnormal COLV expression decreases in later disease stages could explain why skin thickening sometimes improves spontaneously with time. Besides, COLV is correlated to MRSS and disease activity. These findings include COLV as an important regulator of cutaneous thickness in SSc and may add this protein as a new target for future treatments.conferenceObject COLVa2 a Biomarker of Vasculopathy in Scleroderma?(2013) MORAIS, J.; MARTIN, P.; CAMARGO, I. C.; KATAYAMA, M. L.; CARRASCO, S.; GOLDEINSTEIN-SCHAINBERG, C.; PARRAS, E. R.; BARRENCE, F.; VELOSA, A. P.; CAPELOZZI, V. L.; TEODORA, W. R.conferenceObject TRIDIMENSIONAL RECONSTRUCTION, BIOCHEMICAL AND MOLECULAR PROFILE OF COLLAGEN V IN SKIN AND LUNG FIBROBLASTS CULTURE FROM SSc INDICATE A FAILING IN FIBRILOGENESIS(2012) TEODORO, W.; MORAIS, J.; MARTIN, P.; VELOSA, A. P. P.; CARRASCO, S.; SOUZA, R. B. C.; KATAYAMA, M. L.; GOLDEINSTEIN-SCHAINBERG, C.; PARRA, E. R.; CAPELOZZI, V. L.; YOSHINARI, N. H.Background. The type V collagen (COL V) mutations are involved in collagen vascular diseases, such as SSc, in which an unusual accumulation of this collagen was demonstrated (Pathol Res Pract 2004; 200:681). In this context, our purpose was to analyse the 3D reconstruction, biochemical and molecular profile of COL Vα1 and α2 chains in skin and lung fibroblasts culture from patients with SSc. Methods. Lung biopsies of seven patients, skin of six patients and respective matched controls were obtained from SSc according ACR. For fibroblast culture skin and lung evaluations were used the following score: intense expression (1–4), fibroblast number/field (1, 2) and collagen fibres architecture (1–3). The total evaluations were: mild (3–5), moderate (6, 7) and severe (8, 9). COLV 3D reconstruction was performed by confocal microscopy, COL Vα1 and Vα2 gene expression in fibroblasts of skin and lung was performed in PCR–RT and COLV protein expression by immunoblotting. Results. The structure of COL V fibre in 3D reconstruction showed distorted and strongly thickened fibres in skin and lung fibroblasts with irregular bundles of COL V distributed in parallel and perpendicular arrangements resulting in a dense network in SSc patients compared with thin fibres pattern from the healthy controls. Collagen quantification showed increase of COL V fibres expression in SSc cutaneous fibroblast [82.5 (9.5)% vs 47.5 (9.5)%, P = 0.002] and lung fibroblast 38.87 (2.99)% vs 20.33 (7.50)%, P = 0,002) compared with respective controls. The molecular evaluation demonstrated an increased of COL Vα1 and α2 mRNA expression in SSc fibroblast skin when compared with control [1.375 (0.373) au vs 0.0047 (0.0013) au, P = 0.05). Similar results were observed in lung [1.61 (0.654) vs 0.99 (0.51) au; P = 0.05).The proportion COL Vα1/COL Vα2 mRNA in fibroblast lung and skin was higher in SSc than in controls being the chains ratio 1 : 2. COL V chains from skin and lung fibroblasts presented alteration of molecular weight of the quoted chain. Conclusion. The overexpression and the unusual organization of COLV fibres, besides the biochemical changes, suggest an interference with the fibrillogenesis process in skin and pulmonary fibrosis from SSc patients, reinforcing the participation of this collagen in pathogenesis of SSc and open new therapeutic perspectives for these patients.- Ultra High Dilution of triiodothyronine modifies cellular apoptosis in Rana catesbeiana tadpole tail in vitro(2011) GUEDES, J. R. P.; CARRASCO, S.; FERREIRA, C. M.; BONAMIN, L. V.; SOUZA, W.; GOLDENSTEIN-SCHAINBERG, C.; PARRA, E. R.; CAPELOZZI, V. L.Background: Ultra High Dilutions (UHD) are diluted beyond the Avogadro limit with dynamization (dilution with succussion). The process of anuran amphibian metamorphosis is controlled by thyroid hormones, including the resorption of the tadpole tail. Methods: A randomized and blinded study was performed to investigate the influence of triiodothyronine (T3) 5.10(-24) M (10cH) on apoptosis induced by T3 100 nM in Rana catesbeiana tadpoles' tail tips, in vitro. Explants were randomized to three groups: control: no T3 in pharmacological or UHD dose; test: T3 100 nM and challenged with T3 10cH (UHD); positive control: T3 100 nM, treated with unsuccussed ethanol. The apoptotic index and the area of explants of test and control groups at the first and final day of the experiment were compared by t-test. Results: There was no difference in tail tip area between test and control groups, but a significantly higher (p < 0.01) index of apoptosis in explants of the test group. Conclusion: This data suggest that T3 10cH modifies the effect of T3 at pharmacological dose, opening new perspectives for further studies and investigation of the dose effect curve. Homeopathy (2011) 100, 220-227.
conferenceObject Stem cells of adipose rabbit tissue are stimulate into chondrocyte-like phenotype by collagen V in vitro(2012) CRUZ, I. Brindo da; GOLDENSTEIN-SCHAINBERG, C.; FULLER, R.; VELOSA, A. P. P.; CARRASCO, S.; PARRA, E. R.; CAPELOZZI, V. L.; YOSHINARI, N. H.; TEODORO, W. R.Introduction: Among a variety of biological functions, including an anti-inflammatory effect, collagen V (COLV) regulates the diameter of collagen fibers with an important role in the development of functional tissues. Therefore the aim of this study was to evaluate, in rabbits, the influence of COLV in the induction of differentiation of adipose tissue-derived stem cells to a chondrocyte-like cell phenotype. Materials and Methods: New Zealand Rabbits were used as source of adipose tissues for the isolation of mesenchymal stem cells (MSCs). Preliminary characterization of mesenchymal lineage and differentiation into chondrocyte-like phenotype was confirmed by immunofluorescence analysis using antibodies to collagens I, II (polyclonals), III and CD34 (monoclonals). After 2 and 3 weeks in culture with and without COLV, the cell aggregates were fixed for 2 h in 4% formaldehyde, dehydrated with ethanol, washed with xylene and embedded in paraffin. Different sections were cut and stained with Toluidine blue, Alcian blue and Picrosirius red respectively. Results: Proteoglicans and collagen fibers were observed by assessment of the different stains, confirming the collagen expression. Remarkably, compared to control cultures, in the presence of COLV timulation, MSCs were capable to increase production of collagen I and II, confirming its chondrocyte-like cell phenotype. Conclusion: We conclude that COLV may facilitate the differentiation of rabbit adipose tissue-derived stem cells into a chondrocyte-like phenotype and this result can be considered to be candidate for therapies.conferenceObject COLVa2 a Biomarker of Vasculopathy in Scleroderma?(2013) MORAIS, J.; MARTIN, P.; CAMARGO, I. C.; KATAYAMA, M. L.; CARRASCO, S.; GOLDEINSTEIN-SCHAINBERG, C.; PARRAS, E. R.; BARRENCE, F.; VELOSA, A. P.; CAPELOZZI, V. L.; TEODORA, W. R.conferenceObject COLLAGEN V AND DECORIN INTERACTION ARE INVOLVED IN PULMONARY FIBROSIS OF SSc(2012) TEODORO, W.; MARCELINO, A.; CRUZ, I. C. Brindo da; VELOSA, A. P. P.; MARTIN, P.; CARRASCO, S.; GOLDENSTEIN-SCHAINBERG, C.; PARRA, E. R.; CAPELOZZI, V. L.; YOSHINARI, N. H.Aims. Type V collagen (COL V) is involved in SSc pathogenesis since immunization of health rabbits with this protein induces an experimental model reproducing the main pathogenic manifestation of this disease. We have demonstrated an increased amount of unusual COL V fibrils deposition in lung of SSc patients indicating an important role for this protein in fibrosis. Formation of fibrotic tissue can be induced both by cytokines and cell–matrix interaction involving signalization mechanism. COL V and decorin participate of this mechanism interfering with fibrilogenesis. The aim was to evaluate COL V and decorin expression in pulmonary tissue and to characterize biochemical profile of COLV from lung fibroblasts culture from SSc patients. Methods. We evaluated COL V and decorin expression as well as 3D reconstruction using IF in lung specimens from six patients with SSc without pulmonary hypertension and six normal individuals died from trauma. The amount of COL V in lung sections was evaluated with software Image Pro-Plus 6.0 in Olympus-BX51. Quantitative immunoblotting was used to characterize COL V biochemistry from lung fibroblasts culture. Results. It was found that the structure of COL V fibres was distorted and strongly thickened in lung tissue from SSc patients compared with thin fibres pattern in the healthy controls. Decorin was distributed around COL V fibrils in the bronchovascular interstitium and vascular walls. Histomorphometric analysis of SSc lung demonstrated increased expression of both COL V and decorin when compared with the control (68.52 + 7.36% vs 5.01 + 2.12%, P < 0.01 and 22.99 + 0.59% vs 32.93 + 3.81%, P = 0.01, respectively). The semiquantitative immunoblotting detected an increased high molecular weight COL V fraction in patients when compared with the controls (P = 0.02). Conclusions. The overexpression and unusual organization of COL V fibres with biochemical changes associated to increased decorin indicates that matrix signalization pathway is involved in COL V fibrillogenesis process in SSc pulmonary fibrosis.