Department of Endocrinology, The First Affiliated Hospital of Bengbu Medical College, No. 287, Changhuai Road, Bengbu, 233004, Anhui, China.
Department of Endocrinology Tongren Hospital Affiliated to Jiaotong University, No. 1111, Xianxia Road, Changning District, Shanghai, 200336, China.
Appl Biochem Biotechnol. 2024 Jul;196(7):3731-3746. doi: 10.1007/s12010-023-04721-5. Epub 2023 Sep 28.
Diabetic nephropathy (DN) is a common clinical syndrome in diabetic patients. Functional characterization of non-coding (ncRNAs) involved in the progression of DN can provide insights into the diagnosis and therapeutic management of DN. Human kidney proximal tubular epithelial cells (HK-2) were challenged by high glucose (HG, 50 mM) as a cell model of DN. The expression level of long non-coding RNA (lncRNA) ZFAS1 was quantified by qRT-PCR. The proteins and cytokines related to fibrosis and scortosis in DN (NLRP3, GSDMD-N, IL-1β and Caspase 1, fibronectin, collagen I, collagen III, IL-1β, and IL-18) were examined by western blot or ELISA. RNA precipitation and luciferase reporter activity experiments were conducted to assess the molecular associations. ZFAS1 and SGK1 were highly induced in HK-2 cells challenged with HG, while miR-525-5p downregulated upon HG treatment. ZFAS1 knockdown attenuated HG-induced fibrosis and scortosis in HK-2 cells by reducing the levels of NLRP3, GSDMD-N, Caspase 1, fibronectin, collagen I/III, IL-1β, and IL-18. Mechanically, ZFAS1 knockdown protected HK-2 cells from HG-induced injury by upregulating miR-525-5p and repressing SGK1 expression. Overall, our results suggest that knocking down ZFAS1 may be formulated as a protective strategy in ameliorating DN progression through regulating miR-525-5p/SGK1 pathway. Targeting ZFAS1 could be further explored as a potential approach for the management of DN.
糖尿病肾病（DN）是糖尿病患者常见的临床综合征。研究参与 DN 进展的非编码（ncRNAs）的功能特征，可以深入了解 DN 的诊断和治疗管理。高浓度葡萄糖（HG，50mM）刺激人肾近端小管上皮细胞（HK-2）作为 DN 的细胞模型。通过 qRT-PCR 定量检测长链非编码 RNA（lncRNA）ZFAS1 的表达水平。通过 Western blot 或 ELISA 检测与纤维化和硬化相关的蛋白质和细胞因子（DN 中的 NLRP3、GSDMD-N、IL-1β 和 Caspase 1、纤连蛋白、I 型胶原、III 型胶原、IL-1β 和 IL-18）。进行 RNA 沉淀和荧光素酶报告基因活性实验以评估分子相关性。在 HG 刺激的 HK-2 细胞中，ZFAS1 和 SGK1 高度诱导，而 miR-525-5p 在 HG 处理后下调。ZFAS1 敲低通过降低 NLRP3、GSDMD-N、Caspase 1、纤连蛋白、I/III 型胶原、IL-1β 和 IL-18 的水平，减弱了 HG 诱导的 HK-2 细胞纤维化和硬化。机制上，ZFAS1 敲低通过上调 miR-525-5p 和抑制 SGK1 表达来保护 HK-2 细胞免受 HG 诱导的损伤。总之，我们的研究结果表明，敲低 ZFAS1 可能通过调节 miR-525-5p/SGK1 通路，作为改善 DN 进展的保护策略。靶向 ZFAS1 可能进一步被探索作为治疗 DN 的潜在方法。
