Effects Of Methylglyoxal On Oxidative Stress And Cytokine Profiles In Jurkat Cells

Objective: Recent observations have been proposed to explain the hyperglycosemia -induced pathogenesis of vascular complication of diabetes. Increased formation of the very reactive dicarbonyl compound MGO, one of the side-product of glycolysis, and MGO-drived AGEs seem to be implicated in the development of diabetic vascular complication. The researchs show that MGO can accelerate the development of atherosclerosis,closely relate with T lymphocyte immune dysfunction.We therefore examined whether MGO was capable of inducing in Jurkat cells to address the possible mechanism by which MGO accelerated pathologic change in diabetes.Methods:①Jurkat cells which prestimulated by PHA were incubated with 0-60μM concentration of MGO for 24h. The cells viability was detected by MTT. The same Jurkat cells were incubated with 0-60μM MGO for 8h, followed by determination of ROS and MnSOD activity. Total and phosphorylated p38,JNK in extracts were assessed using western-blot after treatment of 30μM MGO for 0-60min. After 24h incubation with 0-60μM MGO, the medium was collected and the expression of TNF-αand IFN-γwere tested by ELISA.②To further explore the relationship between MGO and inflammatory response, various pharmacologlical agents were supplemented to the incubation medium including p38,JNK inhibitor and NAC,AG.Results: No changes in cells viability was observed when Jurkat cells were treated with 0-60μM MGO for 24h(p>0.05).Incubation of Jurkat cells with 15-60μM MGO significantly induced ROS production compared with that of untreated cells(p<0.01). Coincubation with NAC and AG decreased MGO-induced ROS production.Decreased of MnSOD activities also was observed in Jurkat cells with MGO treatmen(tp<0.01), and it was reversed by the addition of NAC and AG. Incubation of Jurkat cells with 30μM MGO results in significant increase in p38,JNK phosphorylation(p<0.01). In addition 15-60μM MGO significantly increased production of TNF-αand IFN-γby Jurkat cells in concentration dependent manne(rp<0.01). NAC or p38,JNK inhibitors reversed the secretion of proinflammatory cytokines.Conclusion: These data demonstrate that MGO induced the release of TNF-αand IFN-γin Jurkat cells via oxidative stress,p38,JNK signaling pathway.