Department of Nephrology, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou City, China .
J Cardiovasc Pharmacol. 2022 Jun 1;79(6):887-895. doi: 10.1097/FJC.0000000000001262.
Circular RNAs (circRNAs) have been verified as vital regulators in various diseases, including membranous nephropathy (MN). Therefore, the role of circ_CDYL in podocyte apoptosis and MN was investigated. The real-time quantitative polymerase chain reaction was performed to measure the expression of circ_CDYL, microRNA-149-5p (miR-149-5p), and tumor necrosis factor superfamily member 11 (TNFSF11) in podocytes. In addition, angiotensin II (Ang II) was used to induce apoptosis of podocytes. The apoptosis-related protein expression was quantified by western blot assay. The apoptosis of podocytes was evaluated by flow cytometry assay. The interaction relationship between miR-149-5p and circ_CDYL or TNFSF11 was confirmed by dual-luciferase reporter assay. Circ_CDYL was significantly overexpressed in MN patients and Ang II-induced podocytes compared with control groups. Importantly, loss-of-functional experiments indicated that knockdown of circ_CDYL protected podocytes from Ang II-induced apoptosis. MiR-149-5p was verified as target of circ_CDYL and negatively correlated with circ_CDYL expression in MN patients. Knockdown of circ_CDYL-mediated effects on Ang II-induced podocyte cells were abolished by silencing miR-149-5p. Besides, the upregulation of miR-149-5p could suppress apoptosis in Ang II-induced podocyte cells by targeting TNFSF11. Under Ang II stimulation, the upregulation of TNFSF11 could increase the expression of TNFSF11 and induce apoptosis in circ_CDYL-silencing podocytes. Our results confirmed that circ_CDYL specifically targeted miR-149-5p/TNFSF11 pathway to regulate Ang II-induced apoptosis in podocytes, which might be useful diagnostic biomarkers in MN.
环状 RNA（circRNAs）已被证实为多种疾病的重要调控因子，包括膜性肾病（MN）。因此，研究了 circ_CDYL 在足细胞凋亡和 MN 中的作用。采用实时定量聚合酶链反应检测足细胞中 circ_CDYL、微小 RNA-149-5p（miR-149-5p）和肿瘤坏死因子超家族成员 11（TNFSF11）的表达。此外，用血管紧张素 II（Ang II）诱导足细胞凋亡。通过 Western blot 检测法定量测定凋亡相关蛋白的表达。通过流式细胞术检测足细胞的凋亡。通过双荧光素酶报告基因检测法证实 miR-149-5p 与 circ_CDYL 或 TNFSF11 的相互作用关系。与对照组相比，MN 患者和 Ang II 诱导的足细胞中 circ_CDYL 的表达显著上调。重要的是，功能丧失实验表明，circ_CDYL 的敲低可保护足细胞免受 Ang II 诱导的凋亡。miR-149-5p 被证实为 circ_CDYL 的靶基因，与 MN 患者中 circ_CDYL 的表达呈负相关。沉默 miR-149-5p 可消除 circ_CDYL 介导的对 Ang II 诱导的足细胞的作用。此外，miR-149-5p 的上调可通过靶向 TNFSF11 抑制 Ang II 诱导的足细胞凋亡。在 Ang II 刺激下，TNFSF11 的上调可增加 TNFSF11 的表达并诱导 circ_CDYL 沉默的足细胞凋亡。我们的研究结果证实，circ_CDYL 特异性靶向 miR-149-5p/TNFSF11 通路调节 Ang II 诱导的足细胞凋亡，这可能对 MN 的诊断具有重要意义。
