Methylglyoxal and Advanced Glycation End-Products Promote Cytokines Expression in Peritoneal Mesothelial Cells Via MAPK Signaling

doi:10.1097/MAJ.0000000000000394

The American Journal of the Medical Sciences

Volume 349, Issue 2, February 2015, Pages 105-109

https://doi.org/10.1097/MAJ.0000000000000394 Get rights and content

Abstract

Background

Peritoneal dialysis fluid degrades glucose into glucose degradation products that impair peritoneal mesothelial cell functions. These compounds are known to interfere with many cellular functions and to promote the formation of advanced glycation end products (AGEs). This study aimed to investigate the biological effects and the underlying mechanism of glucose degradation products and AGEs on mesothelial cells.

Methods

Cell proliferation was determined using [3H]-thymidine incorporation assay. Real-time polymerase chain reaction and enzyme linked immunosorbent assay (ELisA) were used to determine the mRNA and protein expression of cytokines. Reactive oxygen species production in mesothelial cells was determined by flow cytometry. Western blot was used to measure the protein expression of p38 MAPK.

Results

Methylglyoxal (MGO) and AGE-human serum albumin (AGE-HsA) inhibited human peritoneal mesothelial cells proliferation in a dose- and time-dependent manner. The mRNA and protein expression of cytokines including vascular endothelial growth factor (VEGF) and monocyte chemotactic protein-1 (MCP-1) was significantly increased after treatment with MGO and AGE-HSA. Also, the antioxidant N-acetylcysteine (NAC) inhibited MGO- or AGE-HSA-induced reactive oxygen species generation. Western blot showed that MGO and AGE increased the phosphorylation levels of p38 MAPK, which was significantly attenuated after treatment of NAC or p38 MAPK inhibitor SB203580. Furthermore, AGE- or MGO-induced increased expression of VEGF and MCP-1 was significantly reduced in the presence of NAC or SB203580.

Conclusions

Together, this study suggested that AGE or MGO promoted VEGF and MCP-1 expression through activation of p38 MAPK signaling.

Section snippets

Reagents

MGO was purchased from Sigma (St. Louis, MO). AGE-human serum albumin (AGE-HSA) was prepared by incubation of HSA (50 mg/mL) with D-glucose (0.5 mol/L) in 0.1 mol/L of phosphate buffer (pH 7.4) at 37°C in the dark.¹⁴ AGE-specific fluorescence was determined using ratio spectrofluorometry. AGE-HSA contained 68.5 U/mg protein of AGEs, whereas native albumin contained 0.9 U/mg protein of AGEs. Aliquots were removed after 9 weeks, dialyzed against phosphate-buffered saline and stored frozen at −

MGO and AGE-HSA Inhibited the Proliferation of Mesothelial Cells

To determine the effect of MGO and AGE-HSA on cells proliferation, HMrSV5 cells were subjected to different concentrations of MGO and AGE-HSA, and cell proliferation was analyzed after 24 or 48 hours using [3H]-thymidine incorporation. Results showed that mesothelial cells demonstrated a dose- and time-dependent decrease in cell proliferation after MGO or AGE-HSA treatment compared with the control group (Figures 1A and 1B). MTT assay showed that MGO and AGE-HSA suppressed cell viability in a

DISCUSSION

Long-term PD will result in peritoneal thickening with functional decline. GDPs in the PD fluid may be a cause of peritoneal thickening. During PD, GDPs-formed AGE may play an important role in the peritoneal membrane thickening.¹⁵ In this study, we found that AGE or MGO treatment could enhance the expression of VEGF and MCP-1 through activation of p38 MAPK signaling.

The [3H]-thymidine incorporation assay showed that MGO or AGE-HSA treatment decreased the human peritoneal mesothelial cells

CONCLUSIONS

In summary, we demonstrated that MGO and AGEs enhanced the expression of VEGF and MCP-1 in human peritoneal mesothelial cells through p38 MAPK signaling.

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: The authors have no financial or other conflicts of interest to disclose.

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Clinical InvestigationMethylglyoxal and Advanced Glycation End-Products Promote Cytokines Expression in Peritoneal Mesothelial Cells Via MAPK Signaling

Abstract

Background

Methods

Results

Conclusions

Section snippets

Reagents

MGO and AGE-HSA Inhibited the Proliferation of Mesothelial Cells

DISCUSSION

CONCLUSIONS

Am J Kidney Dis

FEBS Lett

Am J Pathol

Lab Invest

Exp Cell Res

Kidney Int

J Lab Clin Med

Kidney Int

Mol Cell Endocrinol

Outcome of peritoneal dialysis in cirrhotic patients with end-stage renal disease: a 24-years' experience in Taiwan

Clin Nephrol