School of Food Sciences and Engineering, South China University of Technology, Guangzhou 510641, China.
Department of Microbiology, Immunology and Cancer Biology, University of Virginia, Charlottesville, USA.
Free Radic Biol Med. 2019 May 20;136:135-145. doi: 10.1016/j.freeradbiomed.2019.04.007. Epub 2019 Apr 10.
Hyperuricemia is a metabolic disease caused by disorders of purine metabolism, the prevalence of which has increased worldwide. Here, a cell model for high uric acid production was established in vitro employing cultured human kidney cells (HK-2 cells), and its molecular basis was analyzed using gene expression profile. High performance liquid chromatography (HPLC) was used to monitor the content of metabolites in cell culture media. Adenosine addition was found to induce HK-2 cells to produce uric acid precursors (inosine and hypoxanthine). Furthermore, the cell model was verified by confirming the antihyperuricemic effect of the widely used antihyperuricemic drugs allopurinol, probenecid, and febuxostat, as well as reported bioactive peptides and amino acids, encompassing glutathione, tryptophan and carnosine, which significantly reduced uric acid production in the HK-2 cells (p < 0.05). RNA-Seq technology was used to perform a wide transcriptome analysis of the hyperuricemic cell model, and the results demonstrated that it has the potential to be used as a rapid and valid in vitro model to screen antihyperuricemic compounds that mimics in vivo cell growth patterns.
高尿酸血症是一种嘌呤代谢紊乱引起的代谢性疾病，其患病率在全球范围内呈上升趋势。在这里，我们通过体外培养人肾细胞（HK-2 细胞）建立了一种高尿酸产生的细胞模型，并通过基因表达谱分析其分子基础。采用高效液相色谱（HPLC）监测细胞培养物中代谢物的含量。结果发现，腺嘌呤的添加诱导 HK-2 细胞产生尿酸前体（肌苷和次黄嘌呤）。此外，通过验证广泛应用的降尿酸药物别嘌醇、丙磺舒和非布司他以及报道的生物活性肽和氨基酸（包括谷胱甘肽、色氨酸和肌肽）对 HK-2 细胞的降尿酸作用，验证了该细胞模型的有效性，这些物质可显著降低 HK-2 细胞中的尿酸生成（p<0.05）。采用 RNA-Seq 技术对高尿酸血症细胞模型进行了广泛的转录组分析，结果表明，该模型有望成为一种快速有效的体外模型，用于筛选模拟体内细胞生长模式的抗高尿酸化合物。
