Korean J Nephrol.  2001 May;20(3):362-374.

Effect of High Glucose on the Expression of MCP-1 and VCAM-1 in Cultured Human Peritoneal Mesothelial Cells

• Lee JS
• Kim SB
• Lee SK
• Yu SH
• Park JS

Abstract

BACKGROUND: High glucose in peritoneal dialysis solution has been implicated in the pathogenesis of peritoneal fibrosis. Macrophages in peritoneal cavity seem to participate in the process of peritoneal fibrosis through the production of various cytokines and growth factors. Monocyte chemoattractant protein-1(MCP-1) plays a key role in the recruitment of monocytes toward the peritoneal cavity. Vascular cell adhesion molecule-1(VCAM-1) is assumed to be important in the transmigration of monocytes. MCP-1 and VCAM-1 can be induced by various cytokines and growth factors in human peritoneal mesothelial cells(HPMC). However, effect of high glucose on the expression of MCP-1 and VCAM-1 in HPMC has not been known well.
METHODS
Cultured HPMC were conditioned with glucose(5-100mM) or mannitol for varying periods up to 7 days. Cell proliferation and mRNA expression of MCP-1 and VCAM-1 were assessed by MTT assay and Northern blot analysis respectively. MCP-1 protein was measured using ELISA. Chemotactic activity of high glucose-conditioned culture supernant were evaluated by chemotactic assay. Effect of protein tyrosine kinase(PTK) inhibitor on the high glucose-induced MCP-1 mRNA expression was examined.
RESULTS
Glucose inhibited the cell proliferation in a time and dose dependent manner. Northern blot analysis showed that high glucose increased the MCP-1 mRNA expression in a time(2-7days) and dose(15-100mM) dependent manner, but not VCAM-1 mRNA expression. MCP-1 protein in cell culture supernant was also increased. Equivalent osmotic concentration of mannitol had no significant effect. High glucose-conditioned supernant had an increased chemotactic activity for monocyte, which was neutralized by specific anti-MCP-1 antibody. PTK inhibitors such as genistein and herbimycin A suppressed the high glucose-induced MCP-1 mRNA expression in a dose dependent manner.
CONCLUSION
High glucose induced MCP-1 expression in HPMC partly via pathways involving PTK.

Keyword

High glucose; Human peritoneal mesothelial cell; MCP-1; VCAM-1; Protein tyrosine kinase

MeSH Terms

Blotting, Northern
Cell Adhesion
Cell Culture Techniques
Cell Proliferation
Cytokines
Enzyme-Linked Immunosorbent Assay
Genistein
Glucose*
Humans*
Intercellular Signaling Peptides and Proteins
Macrophages
Mannitol
Monocytes
Peritoneal Cavity
Peritoneal Dialysis
Peritoneal Fibrosis
Protein-Tyrosine Kinases
RNA, Messenger
Tyrosine
Vascular Cell Adhesion Molecule-1*
Cytokines
Genistein
Glucose
Intercellular Signaling Peptides and Proteins
Mannitol
Protein-Tyrosine Kinases
RNA, Messenger
Tyrosine
Vascular Cell Adhesion Molecule-1