Radiation-induced alteration of rat mesangial cell transforming growth factor-beta and expression of the genes associated with the extracellular matrix.
We hypothesized that the altered mesangial cell phenotype observed in radiation nephropathy reflects, at least partly, radiation-induced changes in expression of the genes associated with transforming growth factor-beta (TGF-beta) and extracellular matrix (ECM). To test this hypothesis, rat mesangial cells were used between passages 7 and 11 after primary isolation from glomeruli. Cells were placed in serum-free medium 24 h prior to irradiation and irradiated with single doses of 5-20 Gy 137Cs gamma rays; control cells received sham irradiation. After irradiation, the cells were maintained in serum-free medium for up to 48 h postirradiation. Total RNA was isolated, and Northern analysis was performed using cDNA probes for TGF-beta 1, beta 2 and beta 3 and several ECM genes. Irradiation resulted in isoform-specific alterations in TGF-beta mRNA; TGF-beta 1 levels showed a dose-independent increase 24-48 h postirradiation; TGF-beta 3 mRNA levels showed a progressive dose-independent decrease over the same period, decreasing to levels approximately 25% of those seen in controls. These changes were associated with a concomitant increase in levels of mRNA expressed by genes for the components of the ECM; no changes were observed in TGF-beta 2, collagen I, collagen III or decorin. Thus radiation can alter mesangial cell TGF-beta and the expression of the genes involved in ECM, although the nature of this alteration varies for the TGF-beta isoforms and specific ECM genes.
Department of Radiology, University of Iowa, Iowa City 52242, USA.
J
J
Wang
M E
ME
Robbins
D000818:Animals; D058578:Biglycan; D002478:Cells, Cultured; D016326:Extracellular Matrix Proteins / Q000235:genetics*; D005353:Fibronectins / Q000235:genetics; D005786:Gene Expression Regulation / Q000528:radiation effects*; D005920:Glomerular Mesangium; D011509:Proteoglycans / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D016212:Transforming Growth Factor beta / Q000235:genetics*
C546886:Bgn protein, rat; D058578:Biglycan; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D011509:Proteoglycans; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta