Zhao Yingdan, Tang Yunhai, Wang Qingqing, Wu Xia, He Zifan, He Yayun, Tang Zhihuan
Department of Nephrology, Jiading Branch of Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai City, China.
Sci Prog. 2025 Jul-Sep;108(3):368504251372305. doi: 10.1177/00368504251372305. Epub 2025 Aug 28.
ObjectiveThis work aims to elucidate the effect and the regulatory mechanisms of miR-205-5p on podocyte injury and oxidative stress in diabetic nephropathy.MethodsA mouse model of diabetic nephropathy was established. Fasting blood glucose, 24 hours urinary albumin, serum creatinine and blood urea nitrogen of mice were detected. H&E and Tunel staining of mice renal tissues were executed to detect histological changes and apoptosis. A cell model of diabetic nephropathy was constructed by inducing mouse podocytes with high glucose. The function of miR-205-5p on viability, apoptosis, and levels of malondialdehyde, superoxide dismutase and glutathione in the diabetic nephropathy cell model was evaluated by CCK-8 assay, Tunel staining and enzyme-linked immunosorbent assay. Binding of miR-205-5p and vascular endothelial growth factor A was verified by dual luciferase reporter gene assay. Rescue experiment was implemented on the diabetic nephropathy cell model to research whether miR-205-5p regulated diabetic nephropathy development by targeting vascular endothelial growth factor A. Quantitative reverse transcription-polymerase chain reaction and Western blot were for the detection of gene expression.ResultsThe increased fasting blood glucose, 24 hours urinary albumin, serum creatinine and blood urea nitrogen levels, the intensified apoptosis and injury, and the down-regulated miR-205-5p were observed in renal tissues. miR-205-5p relieved podocyte injury in diabetic nephropathy, as it increased cell viability, decreased cell apoptosis, reduced malondialdehyde, and elevated superoxide dismutase and glutathione in the diabetic nephropathy cell model. Vascular endothelial growth factor A was up-regulated in renal tissues of diabetic nephropathy mice, and directly suppressed by miR-205-5p. Vascular endothelial growth factor A up-regulation abolished the protection of miR-205-5p on the diabetic nephropathy cell model.ConclusionsmiR-205-5p might relieve podocyte injury in diabetic nephropathy by suppressing Vascular endothelial growth factor A. It might be a promising target for diabetic nephropathy treatment.
目的
本研究旨在阐明miR-205-5p对糖尿病肾病中足细胞损伤和氧化应激的影响及其调控机制。
方法
建立糖尿病肾病小鼠模型。检测小鼠空腹血糖、24小时尿白蛋白、血清肌酐和血尿素氮。对小鼠肾组织进行苏木精-伊红（H&E）染色和TUNEL染色，以检测组织学变化和细胞凋亡。通过高糖诱导小鼠足细胞构建糖尿病肾病细胞模型。采用CCK-8法、TUNEL染色和酶联免疫吸附测定法评估miR-205-5p对糖尿病肾病细胞模型中细胞活力、细胞凋亡以及丙二醛、超氧化物歧化酶和谷胱甘肽水平的影响。通过双荧光素酶报告基因测定法验证miR-205-5p与血管内皮生长因子A的结合。对糖尿病肾病细胞模型进行挽救实验，以研究miR-205-5p是否通过靶向血管内皮生长因子A来调节糖尿病肾病的发展。采用定量逆转录-聚合酶链反应和蛋白质免疫印迹法检测基因表达。
结果
在肾组织中观察到空腹血糖、24小时尿白蛋白、血清肌酐和血尿素氮水平升高，细胞凋亡和损伤加剧，以及miR-205-5p表达下调。miR-205-5p减轻了糖尿病肾病中的足细胞损伤，因为它提高了糖尿病肾病细胞模型中的细胞活力，减少了细胞凋亡，降低了丙二醛水平，并提高了超氧化物歧化酶和谷胱甘肽水平。血管内皮生长因子A在糖尿病肾病小鼠的肾组织中上调，并受到miR-205-5p的直接抑制。血管内皮生长因子A的上调消除了miR-205-5p对糖尿病肾病细胞模型的保护作用。
结论
miR-205-5p可能通过抑制血管内皮生长因子A来减轻糖尿病肾病中的足细胞损伤。它可能是糖尿病肾病治疗的一个有前景的靶点。
