Establishment of the Cell Model of Hyperuricemia and Its Application in Screening Food-derived Anti-hyperuricemic Peptides
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Abstract:
Hyperuricemia is a metabolic disease caused by purine metabolic disorders that lead to the overproduction and (or) reduced excretion of uric acid, and is an important biochemical basis for gout. In the present study, a cell model of hyperuricemia was established and the anti-hyperuricemic activity of whey protein-derived peptides was studied. Thehepatic (LO2) cells were seeded into a 24-well plate and divided into two groups–normal control and model groups. After 48 h of incubation, the model group was incubated with adenosine-containing medium, while the control group was incubated with fresh basal medium. After further incubation for 36 h, xanthine oxidase was added and the incubation was continued for another 12 h. Finally, the cell culture supernatants were collected and analyzed by high performance liquid chromatography (HPLC) assays. The results demonstrated that the uric acid level of model group was significantly higher than the normal control group, and the optimal adenosine concentration for incubation, cell plating density, and optimal incubation time were 2.5 mmol/L, 105 cell/mL, and 36 h, respectively. Under the best conditions for model construction, febuxostat and whey protein hydrolysate were used to evaluate the anti-hyperuricemic activity of the model, and the results showed that the uric acid content of febuxostat group was markedly decreased in a dose-dependent manner, further suggesting that the hyperuricemia model was successfully established. After hepatic cells were incubated with whey protein hydrolysate, when whey protein-derived proteins reached a certain concentration, the level of uric acid decreased significantly compared with that of model group, indicating that whey protein hydrolysates exhibited anti-hyperuricemic activity.
