Department of General Medicine, Weifang People's Hospital, Weifang, 261041, Shandong, China.
Department of Science and Education, People's Hospital of Gaoxin Weifang, Weifang, 261041, Shandong, China.
Biochem Genet. 2022 Oct;60(5):1471-1487. doi: 10.1007/s10528-021-10146-8. Epub 2021 Nov 3.
Diabetic nephropathy (DN) is the main cause of end-stage renal disease. Circular RNA hsa_circ_0004442 (circTLK1) accelerates the progression of renal cell carcinoma. However, the role of circTLK1 in DN pathogenesis is indistinct. The expression of circTLK1, microRNA-126-5p (miR-126-5p), and microRNA-204-5p (miR-204-5p) was tested by quantitative real-time polymerase chain reaction. The levels of interleukin-6 and interleukin-1β were measured by enzyme-linked immunosorbent assay. The levels of reactive oxygen species and malondialdehyde and the activity of superoxide dismutase were determined with corresponding kits. Several protein levels were evaluated with western blotting. The relationship between circTLK1 and miR-126-5p/miR-204-5p was verified by dual-luciferase reporter assay. CircTLK1 was highly expressed in DN patient's serum and high-glucose (HG)-treated human mesangial cells. Functionally, circTLK1 inhibition reduced HG-induced inflammation, oxidative stress, and ECM accumulation in human mesangial cells. CircTLK1 was verified as a sponge for miR-126-5p and miR-204-5p, which were downregulated in DN patient's serum and HG-treated human mesangial cells. Both miR-126-5p and miR-204-5p upregulation decreased inflammation, oxidative stress, and ECM accumulation in HG-treated human mesangial cells and circTLK1 silencing-mediated influence on HG-induced human mesangial cell injury was overturned by miR-126-5p or miR-204-5p inhibition. Moreover, circTLK1 knockdown blocked the AKT/NF-κB pathway by sponging miR-126-5p/miR-204-5p. CircTLK1 downregulation alleviated HG-induced inflammation, oxidative stress, and ECM accumulation through blocking the AKT/NF-κB pathway via sponging miR-126-5p/miR-204-5p, providing a new mechanism to comprehend the pathogenesis of DN.
糖尿病肾病(DN)是终末期肾病的主要原因。环状 RNA hsa_circ_0004442(circTLK1)可加速肾细胞癌的进展。然而,circTLK1 在 DN 发病机制中的作用尚不清楚。通过实时定量聚合酶链反应检测 circTLK1、微小 RNA-126-5p(miR-126-5p)和微小 RNA-204-5p(miR-204-5p)的表达。通过酶联免疫吸附试验测定白细胞介素-6 和白细胞介素-1β 的水平。用相应的试剂盒测定活性氧和丙二醛的水平以及超氧化物歧化酶的活性。用 Western blot 评估几种蛋白质水平。通过双荧光素酶报告基因实验验证 circTLK1 与 miR-126-5p/miR-204-5p 的关系。DN 患者血清和高糖(HG)处理的人系膜细胞中 circTLK1 表达水平升高。功能上,circTLK1 抑制可减少 HG 诱导的人系膜细胞炎症、氧化应激和细胞外基质积聚。circTLK1 被证实为 miR-126-5p 和 miR-204-5p 的海绵,DN 患者血清和 HG 处理的人系膜细胞中 miR-126-5p 和 miR-204-5p 表达下调。miR-126-5p 和 miR-204-5p 的上调均减少了 HG 处理的人系膜细胞中的炎症、氧化应激和细胞外基质积聚,并且 miR-126-5p 或 miR-204-5p 的抑制可逆转 circTLK1 沉默介导的 HG 诱导的人系膜细胞损伤的影响。此外,circTLK1 敲低通过海绵吸附 miR-126-5p/miR-204-5p 阻断 AKT/NF-κB 通路。circTLK1 下调通过海绵吸附 miR-126-5p/miR-204-5p 阻断 AKT/NF-κB 通路,减轻 HG 诱导的炎症、氧化应激和细胞外基质积聚,为理解 DN 的发病机制提供了新的机制。