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1.
Biochim Biophys Acta ; 1780(12): 1388-94, 2008 Dec.
Article in English | MEDLINE | ID: mdl-18708125

ABSTRACT

An alpha-l-Rhamnose specific lectin site was described on human skin keratinocytes and fibrobasts. The addition of Rhamnose-rich oligo- and polysaccharides (RROPs) to fibroblasts has been shown to stimulate cell proliferation and increase extracellular matrix biosynthesis, suggesting that this lectin site functions as a "true" receptor transmitting messages to the cell interior. It was confirmed here that addition of the Rhamnose-rich polysaccharide, RROP-1, to normal human dermal fibroblasts (NHDFs) and human endothelial cells produced a dose-dependent stimulation of the calcium-signaling pathway, inducing fast and transient increases in Ca2+ influx and intracellular free Ca2+ level. The Rhamnose-rich oligosaccharide RROP-3 as well as l-Rhamnose alone were also able to trigger similar intracellular free Ca2+ concentration increases in NHDFs. Moreover, the recording of the RROP-1-induced modification of the gene-expression profile in fibroblasts showed that this polysaccharide triggered a down-regulation of the expression of several growth factors, adhesion molecules and extracellular matrix proteins involved in pro-tumoral activity and/or fibrotic processes. These results further support the hypothesis of a receptor function for the Rhamnose-recognizing lectin site in fibroblasts. Anti-fibrotic and anti-tumoral potential of RROP-1 remains to be further explored.


Subject(s)
Calcium Signaling , Fibroblasts/metabolism , Gene Expression Profiling , Lectins/metabolism , Rhamnose/metabolism , Skin/cytology , Binding Sites , Calcium/metabolism , Calcium Channels/physiology , Cells, Cultured , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Endothelium, Vascular/cytology , Enzyme-Linked Immunosorbent Assay , Fibroblasts/drug effects , Humans , Intracellular Space/metabolism , Oligonucleotide Array Sequence Analysis , Oligosaccharides/pharmacology , Patch-Clamp Techniques , Polysaccharides, Bacterial/pharmacology , Umbilical Veins/cytology
2.
Biogerontology ; 9(3): 177, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18297408

ABSTRACT

The Maillard reaction and its end products, AGE-s (Advanced Glycation End products) are rightly considered as one of the important mechanisms of post-translational tissue modifications with aging. We studied the effect of two AGE-products prepared by the glycation of lysozyme and of BSA, on the expression profile of a large number of genes potentially involved in the above mentioned effects of AGE-s. The two AGE-products were added to human skin fibroblasts and gene expression profiles investigated using microarrays. Among the large number of genes monitored the expression of 16 genes was modified by each AGE-preparations, half of them only by both of them. Out of these 16 genes, 12 were more strongly affected, again not all the same for both preparations. Both of them upregulated MMP and serpin-expression and downregulated some of the collagen-chain coding genes, as well as the cadherin- and fibronectin genes. The BSA-AGE preparation downregulated 10 of the 12 genes strongly affected, only the serpin-1 and MMP-9 genes were upregulated. The lysozyme-AGE preparation upregulated selectively the genes coding for acid phosphatase (ACP), integrin chain alpha5 (ITGA5) and thrombospondin (THBS) which were unaffected by the BSA-AGE preparation. It was shown previously that the lysozyme-AGE strongly increased the rate of proliferation and also cell death, much more than the BSA-AGE preparation. These differences between these two AGE-preparations tested suggest the possibility of different receptor-mediated transmission pathways activated by these two preparations. Most of the gene-expression modifications are in agreement with biological effects of Maillard products, especially interference with normal tissue structure and increased tissue destruction.


Subject(s)
Gene Expression Profiling , Glycation End Products, Advanced/pharmacology , Skin/drug effects , Cells, Cultured , DNA, Complementary , Fibroblasts/drug effects , Humans , Nucleic Acid Hybridization , Oligonucleotide Array Sequence Analysis , Skin/cytology
3.
Pathol Biol (Paris) ; 54(7): 420-5, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16919895

ABSTRACT

Rhamnose-rich oligo- and polysaccharides (RROPs) were tested for their potential pharmacological properties using human skin fibroblasts in serial cultures. The substances tested were shown to stimulate cell proliferation, decrease elastase-type activity, stimulate collagen biosynthesis, and protect hyaluronan against free radical mediated degradation. These reactions appear to be triggered by the mediation of a specific alpha-L-rhamnose recognizing lectin-site acting as a receptor, transmitting signals to the cell-interior. The rapid increase of intracellular free calcium after addition of RROP-1 and preliminary data using micro arrays appear also to confirm this contention.


Subject(s)
Polysaccharides/chemistry , Polysaccharides/pharmacology , Rhamnose/pharmacology , Adult , Calcium/metabolism , Cell Culture Techniques , Cell Division/drug effects , Collagen/biosynthesis , Female , Fibroblasts/cytology , Fibroblasts/drug effects , Free Radical Scavengers/pharmacology , Humans , Hyaluronic Acid/metabolism , Oligosaccharides/pharmacology , Skin/cytology , Skin/drug effects , Skin Physiological Phenomena
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