Department of Clinical Laboratory, Affiliated Hospital of North Sichuan Medical College, Nanchong, P.R. China.
Department of Laboratory Medicine, North Sichuan Medical College, Nanchong, P.R. China.
Nucleosides Nucleotides Nucleic Acids. 2020;39(8):1162-1178. doi: 10.1080/15257770.2020.1780439. Epub 2020 Jul 9.
The present study aimed to examine the expression of hyperuricemia (HUA)-related factors in the body fluids of HUA patients and in renal tissues and body fluids of HUA mice to elucidate the underlying mechanism of HUA and provide theoretical basis for the diagnosis, prevention and treatment of this disease. A total of 51 HUA patients (HUA group), and 36 healthy subjects (control group) were included in the present study. The peripheral blood and urine were collected from all patients and healthy subjects. A total of 20 male Kunming mice were used to construct HUA model, and another 20 mice were used as controls. The kidney tissues, peripheral blood and urine were collected from all mice. ELISA was performed to determine the levels of interleukin-6 receptor (IL-6R) proteins in the serum and urine of human or mice, while western blotting was employed to determine the protein expression in the kidney tissues of mice. Quantitative real-time polymerase chain reaction was used to measure the expression of mRNA and miR-30b in all sample types. Dual luciferase reporter assay was performed to identify the direct interaction between 3'-untranslated region of IL-6R mRNA and miR-30b. The expression of IL-6R mRNA and protein was increased in serum and urine of HUA patients, while the expression of miR-30b was reduced in HUA patients when compared with healthy subjects. The contents of uric acid, urea nitrogen and creatinine in the blood of HUA mice model were significantly elevated. Similarly, the expression of IL-6R mRNA and protein was increased in kidney, serum and urine of HUA mice model, while the expression of miR-30b was reduced in kidney tissues, serum and urine of HUA mice model. Dual luciferase reporter assay showed that miR-30b was able to bind with 3'-UTR seed region of IL-6R mRNA to regulate its expression. These findings demonstrated that the expression of IL-6R in patients and mouse with HUA is elevated, which is related with the down-regulation of miR-30b. Therefore, miR-30b might participate in the pathological process of HUA by regulating IL-6R.
本研究旨在探讨高尿酸血症(HUA)患者体液和 HUA 小鼠肾组织及体液中与 HUA 相关的因素表达情况,阐明 HUA 的发病机制,为该病的诊断、防治提供理论依据。共纳入 51 例 HUA 患者(HUA 组)和 36 例健康受试者(对照组)。采集所有患者和健康受试者的外周血和尿液。采用 20 只雄性昆明小鼠构建 HUA 模型,另设 20 只小鼠作为对照组。收集所有小鼠的肾组织、外周血和尿液。采用 ELISA 法检测人或小鼠血清和尿液中白细胞介素 6 受体(IL-6R)蛋白水平,采用 Western blot 法检测小鼠肾组织中蛋白表达情况,采用实时荧光定量 PCR 法检测各样本类型中 mRNA 和 miR-30b 的表达情况,采用双荧光素酶报告基因实验鉴定 IL-6R mRNA 3'-UTR 与 miR-30b 之间的直接相互作用。HUA 患者血清和尿液中 IL-6R mRNA 和蛋白表达增加,而 miR-30b 表达降低;HUA 小鼠模型血液中尿酸、尿素氮和肌酐含量明显升高,同时,HUA 小鼠模型肾、血清和尿液中 IL-6R mRNA 和蛋白表达增加,而 miR-30b 表达降低。双荧光素酶报告基因实验表明,miR-30b 可与 IL-6R mRNA 的 3'-UTR 种子区结合,调节其表达。这些结果表明,HUA 患者和小鼠中 IL-6R 的表达升高,与 miR-30b 的下调有关。因此,miR-30b 可能通过调节 IL-6R 参与 HUA 的病理过程。