Fructose induces HK-2 cells to express monocyte chemoattractant protein-1 through uric acid and reactive oxygen species
10.3969/j.issn.1674-8115.2018.04.006
- Author:
Qiao-Ling WANG
1
Author Information
1. Renal Division, Molecular Cell Laboratory for Kidney Diseases, Renji Hospital, Shanghai Jiao Tong University
- Publication Type:Journal Article
- Keywords:
Fructose;
HK-2 cells;
Reactive oxygen species;
Uric acid
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2018;38(4):386-393
- CountryChina
- Language:Chinese
-
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 1000 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, allopurinol 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, 10 and 50 mmol/L, respectively) group. The quantitative PCR method and Western blotting method were used to observe the expression of MCP-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 H2O2 promoted MCP-1 production in HK-2 cells. NAC completely inhibited MCP-1 production 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.
