High glucose solution and spent dialysate stimulate the synthesis of transforming growth factor-beta1 of human peritoneal mesothelial cells: effect of cytokine costimulation.

OBJECTIVE To investigate the effect of high glucose and spent peritoneal dialysate on the transforming growth factor-beta1 (TGFbeta1) synthesis of cultured human peritoneal mesothelial cells (HPMCs) and to examine the effect of costimulation with high glucose or spent dialysate, and cytokines, interleukin-1beta (IL-1beta), and tumor necrosis factor-alpha (TNFalpha) on TGFbeta1 synthesis of HPMCs. DESIGN HPMCs were exposed to different concentrations of glucose (30, 60, and 90 mmol/L) or spent peritoneal dialysate for 48 hours in the absence or presence of IL-1beta (1 ng/mL) and TNFalpha(1 ng/mL).TGFbeta1 mRNA expression was assessed by Northern blot analysis and TGFbeta1 protein release by Western blot analysis and enzyme-linked immunosorbent assay (ELISA). RESULTS Exposure of HPMCs to high glucose conditions (30, 60, and 90 mmol/L of D-glucose) induced 2.3-, 3.6-, and 4.0-fold increases inTGFbeta1 mRNA expression of HPMC with enhancedTGFbeta1 protein synthesis and secretion into the media, whereas there were no significant changes in TGFbeta1 synthesis with equimolar concentrations of D-mannitol. Incubation with spent dialysate also significantly increased TGFbeta1 mRNA expression and protein secretion compared to control media (p < 0.05). Stimulation with IL-1beta (1 ng/mL) or TNFalpha (1 ng/mL) resulted in a significant increase in TGFbeta1 mRNA expression after 48 hours: 2.7 and 2.1 times the control level, respectively. However,TNFalpha-induced increase in TGFbeta1 mRNA expression was not translated intoTGFbeta1 protein secretion, while IL-1beta stimulation induced a significant increase in TGFbeta1 protein secretion as well as TGFbeta1 mRNA expression. Combined stimulation by high glucose or spent dialysate, together with IL-1beta or TNFalpha, showed a greater increase in TGFbeta1 mRNA expression and protein secretion compared to stimulation by high glucose or spent dialysate alone. CONCLUSION Our results clearly show that high glucose solution and spent dialysate themselves might be sufficient to stimulate the production of TGFbeta1 by peritoneal mesothelial cells. In peritoneal dialysis patients, this state of chronic induction of TGFbeta1 is further exacerbated in the presence of peritonitis because of the stimulatory effect of proinflammatory cytokines, resulting in augmented TGFbeta1 synthesis, thus promoting peritoneal fibrosis.