Fructose induces HK-2 ceils to express monocyte chemoattractant protein-1 through uric acid and reactive oxygen species / 上海交通大学学报（医学版）

ABSTRACT

Objective·To investigate the mechanism of fructose-induced monocyte chemoattratant protein-1(MCP-1) production in HK-2 cells.Methods·The HK-2 cells were divided into fructose incubated (1,5 and 10 mmol/L) group,fructose and ketohexo-kinase inhibitor (KHK-IN) coincubation (fructose 5 mmol/L,KHK-IN was 12,100 and 1 000 nmol/L,respectively) group,uric acid incubation (5,15 and 50 mg/dL) group,fructose and allopurinol co-incubation (fructose 5 mmol/L,allopurinol were 0.01,0.1 and 0.5 mmol/L) group,uric acid and allopurinol co-incubation (uric acid 50 mg/dL,allopttrinol respectively 0.01,0.1and 0.5 mmol/L) group,H2O2 incubation (0.1 and 0.3 mmol/L) group,fructose and N-acetylcysteine (NAC) coincubation (fructose 5 mmol/L,NAC respectively 5,10 and 50 mmol/L) group,and uric acid and NAC co-incubation (uric acid 50 mg/dL,NAC was 5,10and 50 mmol/L,respectively) group.The quantitative PCR method and Western blotting method were used to observe the expression ofMCP-1 mRNA and protein.The effects of fructose and uric acid on the production of ROS in HK-2 cells were observed by using a fluorescent probe.Results·Fructose doseand time-dependently induced MCP-1 gene transcription and protein production in HK-2 cells,which could be blocked by the ketohexo-kinase blockers.Exogenous uric acid induced MCP-1 production in HK-2 cells.Allopurinol inhibited fructose,but not exogenous uric acid-induced MCP-1 expression.Both fructose and uric acid induced ROS generation.Incubation with H2O2promoted MCP-1 production in HK-2 cells.NAC completely inhibited MCP-1production induced by fructose and H2O2.Conclusion·Catalyzed by the ketohexo-kinase,fructose resultes the production of MCP-1 through uric acid and reactive oxygen species.