Zhang Juntai, Cai Yan, Qin Yan, Liu Jie, Ding Jie, Xu Mengying, Yang Li, Zheng Yuanxin, Zhang Xi
Department of Nephrology & Immunology, Affiliated Ganmei Hospital of Kunming Medical University, Kunming, Yunnan, China.
Department of Nephrology, The Fifth Affiliated Hospital of Kunming Medical University, Gejiu, Yunnan, China.
Ren Fail. 2025 Dec;47(1):2484632. doi: 10.1080/0886022X.2025.2484632. Epub 2025 Apr 11.
Diabetic kidney disease (DKD), characterized by mesangial fibrosis and renal dysfunction, is a major microvascular complication of diabetes. Studies have shown that miRNAs are closely related to the progression of DKD. Therefore, in this study, we aimed to explore whether miR-1225-3p can regulate Smad ubiquitin regulatory factor 2 (SMURF2)-mediated carbohydrate response element binding protein (ChREBP) ubiquitination through Rho GTPase-activating protein 5 (ARHGAP5) to affect fibrosis in DKD.
DKD mice were established by intraperitoneally injecting streptozocin (STZ), and a DKD cell model was generated by culturing in media supplemented with 25 mmol/L glucose (high glucose, HG). StarBase was used to predict the target binding sites between miR-1225-3p and ARHGAP5, and a dual-luciferase reporter gene assay was used to verify this relationship. Western blotting, RT-qPCR, flow cytometry, immunoprecipitation, ELISAs, HE staining, and Masson staining were used to detect relevant indicators.
ARHGAP5 and SMURF2 expression was decreased, but ChREBP was highly expressed in the renal tissue of DKD mice and HG-induced mouse mesangial cells (MMCs). miR-1225-3p could target and regulate the transcription of ARHGAP5, and an association between ARHGAP5 and SMURF2 was revealed. miR-1225-3p facilitated fibrosis and oxidative stress in MCCs by inhibiting ARHGAP5. In addition, SMURF2 promoted the ubiquitination of HA-ChREBP, and miR-1225-3p facilitated fibrosis and oxidative stress by mediating the ARHGAP5/SMURF2-mediated ubiquitination of ChREBP in MCCs. Furthermore, the miR-1225-3p inhibitor inhibited fibrosis and inflammation in the renal tissues of DKD mice.
miR-1225-3p facilitates fibrosis and oxidative stress by mediating ARHGAP5/SMURF2-mediated ubiquitination of ChREBP.
糖尿病肾病（DKD）以系膜纤维化和肾功能不全为特征，是糖尿病的一种主要微血管并发症。研究表明，微小RNA（miRNA）与DKD的进展密切相关。因此，在本研究中，我们旨在探讨miR-1225-3p是否能通过Rho鸟嘌呤核苷酸酶激活蛋白5（ARHGAP5）调节Smad泛素调节因子2（SMURF2）介导的碳水化合物反应元件结合蛋白（ChREBP）泛素化，从而影响DKD中的纤维化。
通过腹腔注射链脲佐菌素（STZ）建立DKD小鼠模型，并通过在补充有25 mmol/L葡萄糖（高糖，HG）的培养基中培养建立DKD细胞模型。使用StarBase预测miR-1225-3p与ARHGAP5之间的靶标结合位点，并使用双荧光素酶报告基因测定法验证这种关系。采用蛋白质免疫印迹法、逆转录-定量聚合酶链反应、流式细胞术、免疫沉淀、酶联免疫吸附测定、苏木精-伊红染色和Masson染色检测相关指标。
ARHGAP5和SMURF2的表达降低，但ChREBP在DKD小鼠肾组织和HG诱导的小鼠系膜细胞（MMC）中高表达。miR-1225-3p可以靶向并调节ARHGAP5的转录，并揭示了ARHGAP5与SMURF2之间的关联。miR-1225-3p通过抑制ARHGAP5促进MMC中的纤维化和氧化应激。此外，SMURF2促进HA-ChREBP的泛素化，并且miR-1225-3p通过介导MMC中ARHGAP5/SMURF2介导的ChREBP泛素化促进纤维化和氧化应激。此外，miR-1225-3p抑制剂抑制DKD小鼠肾组织中的纤维化和炎症。
miR-1225-3p通过介导ARHGAP5/SMURF2介导的ChREBP泛素化促进纤维化和氧化应激。
