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Fructose induces HK-2 cells to express monocyte chemoattractant protein-1 through uric acid andreactive oxygen species
WANG Qiao-ling1*, CHEN Xiao-huan2*, NI Zhao-hui1, GU Le-yi1, XU Chen-qi1, DAI Hui-li1
2018, 38 (4):
386.
doi: 10.3969/j.issn.1674-8115.2018.04.006
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, fructoseand 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 50mg/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, 10and 50 mmol/L, respectively) group. The quantitative PCR method and Western blotting method were used to observe the of MCP-1 mRNA andprotein. The effects of fructose and uric acid on the production of ROS in HK-2 cells were observedusing a fluorescent probe. Results · Fructose dose-and time-dependently induced MCP-1 gene transcription and protein production in HK-2 cells, which could be blockedthe 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 . Both fructose and uric acid induced ROS generation. Incubation with H2O2 promoted MCP-1 production in HK-2 cells. NAC completely inhibited MCP-1production inducedfructose and H2O2. Conclusion · Catalyzedthe ketohexo-kinase, fructose resultes the production of MCP-1 through uric acid and reactive oxygen species.
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