Hou Chuanli, Sha Wangqian, Li Yujuan, Yao Maojin, Ren Jiaoyan
School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China.
The First Affiliated Hospital of Guangzhou Medical University, Guangzhou Institute of Respiratory Disease and China State Key Laboratory of Respiratory Disease, Guangzhou 510182, China.
Food Funct. 2022 Oct 17;13(20):10546-10557. doi: 10.1039/d2fo00297c.
Hyperuricemia is a purine metabolism disorder, with increasing prevalence worldwide. Here, a high throughput cell model for screening of antihyperuricemic compounds was set up. Human kidney cells (HK2 cells) were stimulated with adenosine and the resulting supernatant and lysate were then analyzed using high performance liquid chromatography (HPLC). The results showed that hypoxanthine content was increased in both HK2 cells supernatant and xanthine oxidase (XO)-overexpressing HK2 cells lysate, but no uric acid was detected due to lower endogenous XO content in these cells. Exogenous XO was added to the supernatant, and then used to evaluate the antihyperuricemic activity of Febuxostat and two the previously identified peptides, Pro-Gly-Ala-Cys-Ser-Asn (PGACSN) and Trp-Met-Leu (WML). By adding exogenous XO, this combined-adenosine-XO-induced hyperuricemia model was optimized and established, and the Febuxostat and peptides were confirmed to significantly reduce uric acid production in the HK2 cells supernatant ( < 0.05). Therefore, this cell model could be recommended for screening potential bioactive antihyperuricemic compounds.
高尿酸血症是一种嘌呤代谢紊乱疾病，在全球范围内的患病率不断上升。在此，建立了一种用于筛选抗高尿酸血症化合物的高通量细胞模型。用腺苷刺激人肾细胞（HK2细胞），然后使用高效液相色谱法（HPLC）分析所得的上清液和裂解物。结果表明，HK2细胞上清液和过表达黄嘌呤氧化酶（XO）的HK2细胞裂解物中的次黄嘌呤含量均增加，但由于这些细胞中内源性XO含量较低，未检测到尿酸。向上清液中添加外源性XO，然后用于评估非布司他以及两种先前鉴定的肽（脯氨酸 - 甘氨酸 - 丙氨酸 - 半胱氨酸 - 天冬酰胺（PGACSN）和色氨酸 - 甲硫氨酸 - 亮氨酸（WML））的抗高尿酸血症活性。通过添加外源性XO，优化并建立了这种联合腺苷 - XO诱导的高尿酸血症模型，并且证实非布司他和肽可显著降低HK2细胞上清液中的尿酸生成（<0.05）。因此，该细胞模型可推荐用于筛选潜在的具有生物活性的抗高尿酸血症化合物。
