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La proteina de estrés, Heat Shock Cognate (HSC70)se sobreexpresa en osteoblastos sometidos a centrifugación
Overexpression Heat Shock Cognate (HSC70) protein in osteoblasts submited to centrifugal forces
Palabras clave:
proteínas, proteínas de choque térmico HSC70, osteoblastos, electroforesis, inmunocitoquínica, western blotting. (es)proteins, HSC70 heat-shock proteins, osteoblasts, electrophoresis, immunohistochemistry, blotting, Western (en)
Antecedentes. El osteoblasto tiene la capacidad de estimular la neoformación del tejido óseo, mediante la síntesis de proteínas tales como las proteínas morfogenéticas, factores de crecimiento y proteínas colágenas y no colágenas. A su vez, indirectamente controla los procesos de reabsorción sintetizando otra serie de proteínas estimuladoras de la actividad osteoclastogénica.
Objetivo. El propósito de este trabajo de investigación fue analizar la expresión de la proteína de choque térmico HSC70 en un cultivo primario de osteoblastos, después de someterlo a una fuerza tensil mediante centrifugación, determinando el patrón de proteínas mediante análisis electroforético.
Material y métodos. A partir de cráneos de ratones lactantes ICR se obtuvieron cultivos de células con características morfológicas tipo fibroblastoide, con prolongaciones alargadas y citoplasmas ligeramente cuboidales que sugiere un cultivo enriquecido de osteoblastos; los cuales fueron sometidos a centrifugación y se analizó por inmunocitoquímica, electroforesis y Western blot la expresión diferencial de la proteína HSC70.
Resultados.La técnica de electroforesis SDS-PAGE unidimensional no permitió determinar diferencias en el patrón de corrido entre osteoblastos tratados respecto a los no tratados con la fuerza tensil. Sin embargo, la proteína HSC70 se sobre-expresa en osteoblastos sometidos a centrifugación analizada mediante la técnica de Western blot e inmunohistoquímica.
Conclusión.Aunque la electroforesis no determinó diferencias en el patrón de corrido, la proteína de estrés HSC70 se sobre-expresó en osteoblastos sometidos a centrifugación analizada mediante la técnica de Western blot e inmunocitoquímica, sugiriendo que es importante en este tipo de estrés.
Background. Osteoblasts can stimulate new bone formation by protein synthesis, such as, bone morphogenetic proteins, growth factors and collagen and non-collagenous proteins. As well, they controls reabsortion processes, because they can synthesize proteins to promote osteoclastogenic activity, when necessary.
Objetive. The objetive of this work was analyze the expression of the heat shock cognate protein HSC70 in a primary culture of murine osteoblasts, after putting under a tensil force by centrifugation and determining the protein pattern by electrophoretic analysis.
Materials and methods. From skulls of newborn mice ICR, cultures of cells were obtained with morphologic characteristics fibroblasts-like type, extended prolongations and cytoplasms slightly cuboidal and plump, it suggests an enriched culture of osteoblasts.
Results. The technique of unidimensional electroforesis SDS-PAGE did not allow determine differences in the run pattern between osteoblasts treated with tensil force and control cells. Nevertheless, protein HSC70 overexpresion in osteoblasts was confirmed by Western blot and inmunocytochemistry on-are expressed.
Conclusions. Electroforesis did not determine differences between control cell and cell submited to centrifugation. Stress protein HSC70 was expressed in osteoblasts submitted to centrifugación analyzed by Western's blot technology suggesting a important roll of this stres type.
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Derechos de autor 2006 Revista de la Facultad de Medicina
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