Zhou Ju, Li Wu, Chi Xiaowei, Li Dingchun, Yang Chunxia, Duan Zhiwen
Department of Infectious Disease, The First Affiliated Hospital, Kunming Medical University, Kunming 650032, China.
Endocr J. 2025 Jan 6;72(1):79-91. doi: 10.1507/endocrj.EJ24-0008. Epub 2024 Oct 24.
Circular RNAs (circRNAs) play an important role in regulating inflammation and oxidative stress during the pathogenesis of metabolic dysfunction-associated steatotic liver disease (MASLD); however, the underlying mechanism is unclear. This study aimed to determine the role of mmu_circ_0009303 in MASLD. We used a bioinformatics approach to identify potential targets and established an in vitro model of MASLD. Oil red O staining, cell transfection and dual-luciferase reporter assay were used to determine the role of mmu_circ_0009303. The results indicated that the mmu_circ_0009303 expression was significantly increased in the MASLD model both in vitro and in vivo and was associated with oxidative stress levels and inflammation. Moreover, bioinformatics analyses revealed that miRNA-182-5p and Foxo3 are targets of mmu_circ_0009303 and miRNA-182-5p, respectively. In the in vitro MASLD model, mmu_circ_0009303 promoted fat deposition in NCTC1469 cells, which was induced by free fatty acid (FFA) through the regulation of miRNA-182-5p/Foxo3. The expression of miRNA-182-5p and Forkhead box O3 (Foxo3) was associated with mmu_circ_0009303 expression in the liver of mice with MASLD, which was induced by a high-fat diet. Furthermore, mmu_circ_0009303 may be involved in regulating the expression of lipid metabolism-related regulatory proteins, such as CPT1A, SLC27A4, ACBD3, SREBP1, FAS, PPARα, and PPARγ. Taken together, mmu_circ_0009303 promotes oxidative stress, inflammation, and excessive fat accumulation in NCTC1469 cells induced by FFA through the regulation of miRNA-182-5p/Foxo3 and lipid metabolism-related regulatory proteins. These findings provide a potential target for the treatment of MASLD.
环状RNA（circRNAs）在代谢功能障碍相关脂肪性肝病（MASLD）发病机制中调节炎症和氧化应激方面发挥重要作用；然而，其潜在机制尚不清楚。本研究旨在确定mmu_circ_0009303在MASLD中的作用。我们采用生物信息学方法识别潜在靶点，并建立了MASLD的体外模型。通过油红O染色、细胞转染和双荧光素酶报告基因检测来确定mmu_circ_0009303的作用。结果表明，mmu_circ_0009303在体外和体内的MASLD模型中表达均显著增加，且与氧化应激水平和炎症相关。此外，生物信息学分析显示，miRNA-182-5p和Foxo3分别是mmu_circ_0009303和miRNA-182-5p的靶点。在体外MASLD模型中，mmu_circ_0009303通过调节miRNA-182-5p/Foxo3促进游离脂肪酸（FFA）诱导的NCTC1469细胞脂肪沉积。miRNA-182-5p和叉头框O3（Foxo3）的表达与高脂饮食诱导的MASLD小鼠肝脏中mmu_circ_0009303的表达相关。此外，mmu_circ_0009303可能参与调节脂质代谢相关调节蛋白的表达，如肉碱棕榈酰转移酶1A（CPT1A）、溶质载体家族27成员4（SLC27A4）、酰基辅酶A结合结构域蛋白3（ACBD3）、固醇调节元件结合蛋白1（SREBP1）、脂肪酸合酶（FAS）、过氧化物酶体增殖物激活受体α（PPARα）和过氧化物酶体增殖物激活受体γ（PPARγ）。综上所述，mmu_circ_0009303通过调节miRNA-182-5p/Foxo3和脂质代谢相关调节蛋白，促进FFA诱导的NCTC1469细胞氧化应激、炎症和过度脂肪积累。这些发现为MASLD的治疗提供了一个潜在靶点。
