Chen Jie, Zou Libin, Liu Lu, Wu Chunfeng, Hu Mi
Department of Nephrology, Wuhan Third Hospital, Wuhan, Hubei Province, China.
Department of Nephrology, Wuhan Asia General Hospital, Wuhan, Hubei Province, China.
Organogenesis. 2025 Dec;21(1):2519614. doi: 10.1080/15476278.2025.2519614. Epub 2025 Jun 17.
Podocyte damage is a central feature of lupus nephritis (LN), making the identification of potential therapeutic targets to prevent podocyte injury and improve treatment outcomes essential. ORM1 has been suggested as a significant candidate gene in LN. In this study, mouse podocytes were induced using Immunoglobulin G (IgG) extracted from lupus patients. To investigate the role of ORM1, ORM1 knockdown was performed, and the effects on podocyte viability and apoptosis were assessed using the cell counting kit-8 (CCK-8) assay and flow cytometry. Additionally, autophagy markers LC3II/I and p62 were measured by western blotting and immunofluorescence, and the expression of the AMPK/mTOR signaling pathway was evaluated using western blotting. The results showed an upregulation of ORM1 in the LN model. Upon stimulation with IgG from LN patients, ORM1 knockdown reversed the reduction in podocyte viability, decreased the apoptosis rate, and reduced the elevated levels of autophagy, followed by an increase in AMPK phosphorylation and a decrease in mTOR phosphorylation. In conclusion, these results suggest that ORM1 modulates the expression of autophagy-related components in podocytes through the AMPK/mTOR signaling pathway, thereby influencing podocyte damage in the LN model in vitro.
足细胞损伤是狼疮性肾炎（LN）的核心特征，因此确定潜在的治疗靶点以预防足细胞损伤并改善治疗效果至关重要。ORM1已被认为是LN中的一个重要候选基因。在本研究中，使用从狼疮患者中提取的免疫球蛋白G（IgG）诱导小鼠足细胞。为了研究ORM1的作用，进行了ORM1基因敲低，并使用细胞计数试剂盒-8（CCK-8）检测和流式细胞术评估其对足细胞活力和凋亡的影响。此外，通过蛋白质免疫印迹法和免疫荧光法检测自噬标志物LC3II/I和p62，并使用蛋白质免疫印迹法评估AMPK/mTOR信号通路的表达。结果显示LN模型中ORM1上调。在用LN患者的IgG刺激后，ORM1基因敲低逆转了足细胞活力的降低，降低了凋亡率，并降低了自噬水平的升高，随后AMPK磷酸化增加，mTOR磷酸化减少。总之，这些结果表明ORM1通过AMPK/mTOR信号通路调节足细胞中自噬相关成分的表达，从而在体外影响LN模型中的足细胞损伤。
