Effects of nonenzymatic glycosylation of mesangial matrix on proliferation of mesangial cells

Susan T. Crowley, Michael Brownlee, Diane Edelstein, Joseph A. Satriano, Tetsuo Mori, Pravin C. Singhal, Detlef O. Schlondorff

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Abstract

Cross-linking of cell matrix components by nonenzymatic glycosylation may contribute to diabetic glomerulopathy. We examined the effects of modification of matrix by nonenzymatic glycosylation on mesangial cell function. Matrix was generated by growing mesangial cells in tissue culture for 2 wk and removing the cells with a detergent cell-lysis solution. By indirect immunofluorescence and Northern-blot analysis, the remaining matrix contained laminin, fibronectin, and collagens type I and IV. The matrix was modified by incubation for 24 h with 50 mM glycolaldehyde, a highly reactive cross-linking nonenzymatic glycosylation product, or for 2 wk with 200 mM glucose-6-phosphate (G6P). Modification was carried out with or without equimolar aminoguanidine, an inhibitor of cross-link formation. Nonenzymatic glycosylation of the matrix by glycolaldehyde or G6P was confirmed by fluorometry and [14C]G6P incorporation and was prevented by aminoguanidine. [3H]thymidine incorporation for 24 h by mesangial cells plated onto unmodified or modified matrix was then performed. Modification of matrix had no effect on attachment of mesangial cells, determined 4 h after plating. Nonenzymatic glycosylation of matrix by glycolaldehyde or G6P significantly inhibited thymidine incorporation by mesangial cells. This effect was partially reversible by aminoguanidine. Aminoguanidine-modified matrix had no effect on thymidine incorporation. Thymidine-incorporation results were confirmed by direct cell counting. We conclude that modification of matrix by nonenzymatic glycosylation influences growth of mesangial cells, which could contribute to the mesangial abnormalities of diabetic glomerulopathy.

Original languageEnglish (US)
Pages (from-to)540-547
Number of pages8
JournalDiabetes
Volume40
Issue number5
StatePublished - May 1991

Fingerprint

Mesangial Cells
Glycosylation
Glucose-6-Phosphate
Thymidine
Fluorometry
Collagen Type IV
Laminin
Cellular Structures
Indirect Fluorescent Antibody Technique
Collagen Type I
Fibronectins
Northern Blotting
Detergents
pimagedine
Growth
glycolaldehyde

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Internal Medicine

Cite this

Crowley, S. T., Brownlee, M., Edelstein, D., Satriano, J. A., Mori, T., Singhal, P. C., & Schlondorff, D. O. (1991). Effects of nonenzymatic glycosylation of mesangial matrix on proliferation of mesangial cells. Diabetes, 40(5), 540-547.

Effects of nonenzymatic glycosylation of mesangial matrix on proliferation of mesangial cells. / Crowley, Susan T.; Brownlee, Michael; Edelstein, Diane; Satriano, Joseph A.; Mori, Tetsuo; Singhal, Pravin C.; Schlondorff, Detlef O.

In: Diabetes, Vol. 40, No. 5, 05.1991, p. 540-547.

Research output: Contribution to journalArticle

Crowley, ST, Brownlee, M, Edelstein, D, Satriano, JA, Mori, T, Singhal, PC & Schlondorff, DO 1991, 'Effects of nonenzymatic glycosylation of mesangial matrix on proliferation of mesangial cells', Diabetes, vol. 40, no. 5, pp. 540-547.
Crowley ST, Brownlee M, Edelstein D, Satriano JA, Mori T, Singhal PC et al. Effects of nonenzymatic glycosylation of mesangial matrix on proliferation of mesangial cells. Diabetes. 1991 May;40(5):540-547.
Crowley, Susan T. ; Brownlee, Michael ; Edelstein, Diane ; Satriano, Joseph A. ; Mori, Tetsuo ; Singhal, Pravin C. ; Schlondorff, Detlef O. / Effects of nonenzymatic glycosylation of mesangial matrix on proliferation of mesangial cells. In: Diabetes. 1991 ; Vol. 40, No. 5. pp. 540-547.
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