Gene expression of monocyte chemoattractant protein-1 in human monocytes by exposure to advanced glycosylation end products
Abstract
Aim: To explore the effects of advanced glycosylation end products (AGEP) on monocyte chemoattractant protein-1 (MCP-1) gene expression in human peripheral blood monocytes/macrophages (PBMC).
Methods: Expression of MCP-1 mRNA in PBMC incubated with AGEP-bovine serum albumin (AGEP-BSA) was examined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) with beta-actin as an internal standard. Sequencing of RT-PCR products was performed to confirm the specificity of amplification for MCP-1 gene.
Results: AGEP-BSA stimulated monocytes to express MCP-1 mRNA in a glucose-concentration-related fashion. The levels of MCP-1 mRNA were increased slightly when monocytes were exposed to AGEP-BSA 200 mg/L (glycosylated with glucose 20 mmol/L), and increased markedly when exposed to AGEP-BSA 200 mg/L (glycosylated with glucose 50 mmol/L), but decreased slightly when exposed to AGEP-BSA 200 mg/L (glycosylated with glucose 80 mmol/L). Expression of MCP-1 mRNA was undetectable in freshly isolated monocytes, but was induced at 12 h and reached a maximal level at 24 h and was almost undetectable at 36 h after the monocytes were incubated with AGEP-BSA 200 mg/L (P < 0.01).
Conclusion: AGEP enhanced MCP-1 mRNA expression in human PBMC.
Keywords:
Methods: Expression of MCP-1 mRNA in PBMC incubated with AGEP-bovine serum albumin (AGEP-BSA) was examined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) with beta-actin as an internal standard. Sequencing of RT-PCR products was performed to confirm the specificity of amplification for MCP-1 gene.
Results: AGEP-BSA stimulated monocytes to express MCP-1 mRNA in a glucose-concentration-related fashion. The levels of MCP-1 mRNA were increased slightly when monocytes were exposed to AGEP-BSA 200 mg/L (glycosylated with glucose 20 mmol/L), and increased markedly when exposed to AGEP-BSA 200 mg/L (glycosylated with glucose 50 mmol/L), but decreased slightly when exposed to AGEP-BSA 200 mg/L (glycosylated with glucose 80 mmol/L). Expression of MCP-1 mRNA was undetectable in freshly isolated monocytes, but was induced at 12 h and reached a maximal level at 24 h and was almost undetectable at 36 h after the monocytes were incubated with AGEP-BSA 200 mg/L (P < 0.01).
Conclusion: AGEP enhanced MCP-1 mRNA expression in human PBMC.