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1 BY CULTURED HUMAN MESANGIAL CELLS
1 Physiology, Universidad de Alcala, Alcala de Henares, Madrid, Spain
* To whom correspondence should be addressed. E-mail: mpiedad.ruiz{at}uah.es.
Progressive renal diseases are characterized by an increased synthesis of extracellular matrix (ECM) components. The mechanisms involved in the development of these alterations are incompletely known, but a crucial role for TGF-
1 has been suggested. Moreover, the ability of the ECM to modulate the phenotypic expression of different cell types has been widely described. In experiments presented here, human mesangial cells (HMC) were grown on collagen types I (COL I) or IV (COL IV). ECM protein and TGF-1 mRNA expression were evaluated by Northern blot, and TGF-1 secretion by ELISA. The involvement of tyrosine-kinase and serine-threonine kinase pathways were studied by western blot, immunofluorescence and in vitro kinase assays. HMC cultured on COL I showed an increased mRNA expression of collagen types I and IV, fibronectin and TGF-1. Both tyrosine phosphorylation and integrin-linked kinase (ILK) activity increased when HMC were cultured on COL I, and the blockade of these pathways inhibited the increased secretion of TGF-1. In conclusion, the present results support a role for extracellular collagen I in the regulation of TGF-1 synthesis during progressive renal sclerosis and fibrosis and subsequent increase in newly synthesized ECM proteins. Additionally, along with the tyrosine kinases, ILK participates in the genesis of this effect.
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