INTRACELLULAR GLUTATHIONE INFLUENCES COLLAGEN GENERATION BY MESANGIALCELLS

The cellular redox state is altered in a number of pathological condit ions, including various forms of glomerular injury and diabetes. For e xample, glucose, via the pentose phosphate pathway generates NADPH, wh ich maintains glutathione (GSH) (part of a major intracellular reducin g system) in its reduced state. GSH in turn influences the activity of transcription factors on gene expression. We therefore examined wheth er changes in cellular GSH influence total collagen synthesis and mRNA levels for collagen I, collagen IV and TGF-beta in SV-40 transformed mouse mesangial cells (MC) maintained in either 5 or 25 mM glucose med ia. Total intracellular GSH was increased by N-acetylcysteine (NAC; 10 mM) or decreased with the GSH synthesis inhibitor buthionine sulfoxim ine (BSO; 0.2 mM) in MC. NAC increased H-3-proline incorporation into collagenase-sensitive protein while BSO decreased it under both glucos e conditions. The presence of BSO did not reverse the increased collag en synthesis seen in the NAC stimulated cells. Northern blot analysis showed increased mRNA levels for collagen I, collagen IV and TGF-beta in cells grown in high glucose (25 mw). NAC increased the mRNA for all three compounds while BSO alone had no effect on these mRNA levels. H owever, BSO reversed the increased mRNA levels for collagen I, IV and TGF-beta seen in the presence of NAC. These findings suggest that the cellular redox state may influence gene transcription in MC, and may h ave implications in explaining injury-associated alterations of mesang ial matrix generation.