CEBPA promotes hepatic steatosis in metabolic dysfunction-associated fatty liver disease in mice via the SREBP-1c/FASN/ACC axis

doi:10.12122/j.issn.1673-4254.2026.10.11

Abstract

Abstract:

Methods To investigate the role of CCAAT/enhancer-binding protein alpha (CEBPA) in metabolic dysfunction-associated fatty liver disease (MAFLD) and whether CEBPA promotes hepatocellular lipogenesis by regulating sterol regulatory element-binding protein-1c (SREBP-1c). Methods Eight-week-old male C57BL/6J mice were assigned to a control group fed a standard diet or a MAFLD model with high-fat diet (HFD) feeding. Hepatic lipid accumulation and the expression of key molecules in the CEBPA/SREBP-1c pathway were assessed using serum biochemical analysis, Western blotting, and Oil Red O staining. In HepG2 cells, CEBPA and SREBP-1c expression was manipulated by plasmid-mediated overexpression or RNA interference. A sodium palmitate/sodium oleate-induced lipid-overload model was used to mimic lipid overload in vitro. Quantitative PCR, Western blotting, Oil Red O staining, and reciprocal rescue experiments were performed to explore the regulatory relationship between CEBPA and SREBP-1c and their effects on lipogenic markers. Results HFD-fed mice developed marked hepatic steatosis with increased serum TC, TG, LDL-C, and FFA levels, significantly upregulated hepatic protein expression of CEBPA, SREBP-1c, fatty acid synthase (FASN), and acetyl-CoA carboxylase (ACC), and enhanced lipid deposition. In HepG2 cells, SREBP-1c knockdown reduced FASN and ACC expression, whereas SREBP-1c overexpression increased their expression in both basal and lipid-overload conditions. Similarly, CEBPA knockdown suppressed SREBP-1c, FASN, and ACC expression and reduced intracellular lipid accumulation, while CEBPA overexpression produced the opposite effects. Rescue experiments showed that SREBP-1c overexpression partially reversed the inhibitory effects of CEBPA knockdown on FASN and ACC, whereas SREBP-1c silencing attenuated CEBPA-induced upregulation of these lipogenic molecules. Conclusion CEBPA promotes hepatic lipogenesis possibly by regulating the SREBP-1c/FASN/ACC axis in both basal and lipid-overload conditions, suggesting that the CEBPA-SREBP-1c pathway contributes to hepatic steatosis in MAFLD and represents a potential therapeutic target.

Key words: metabolic dysfunction-associated fatty liver disease, CCAAT/enhancer-binding protein alpha, sterol regulatory element-binding protein 1c, fatty acid synthesis, lipid metabolism

Leiying ZHANG, Liyang ZOU, Lingling ZHOU, Xin MA, Xiaomin LIU, Huan YAO, Yuan ZHUANG, Yang YU. CEBPA promotes hepatic steatosis in metabolic dysfunction-associated fatty liver disease in mice via the SREBP-1c/FASN/ACC axis[J]. Journal of Southern Medical University, 2026, 46(10): 2415-2423.

Figures/Tables 4

Fig.1 Effects of CEBPA on SREBP-1c, FASN, and ACC expression and intracellular lipid accumulation in HepG2 cells. A, B: Efficiency of CEBPA knockdown and detected by qPCR and Western blotting. C: Expression levels of SREBP-1c, FASN and ACC after CEBPA knockdown or overexpression detected by qPCR and Western blotting. D: Intracellular lipid accumulation observed using Oil Red O staining. The right panels show enlarged views (Original magnification: ×400) of the red dashed boxes in the left panels (×100). Data are presented as Mean±SD (n=3). *P<0.05, **P<0.01, ***P<0.001 vs control group.

Fig.2 Effects of SREBP-1c on FASN, ACC expression and intracellular lipid accumulation in HepG2 cells. A, B: Knockdown and overexpression efficiency of SREBP-1c detected by qPCR and Western blotting. C: Expression levels of FASN and ACC in HepG2 cells in routine culture conditions detected by qPCR and Western blotting. D: FASN and ACC protein expression in HepG2 cells with high-lipid stimulation detected by Western blotting. E: Intracellular lipid accumulation in HepG2 cells in routine culture and high-lipid conditions observed by Oil Red O staining. The right panels show enlarged views (×400; Scale bar=50 μm) of the red dashed boxes in the left panels (×100; Scale bars=100 μm). Data are presented as Mean±SD (n=3). **P<0.01, ***P<0.001 vs control group.

Fig.3 CEBPA regulates FASN and ACC expression through SREBP-1c. A: FASN and ACC mRNA levels in different treatment groups detected by qPCR. B: FASN and ACC protein levels detected by Western blotting. C: Densitometric analysis of Western blot bands. Data are presented as Mean±SD (n=3). **P<0.01, ***P<0.001 vs control group.

Fig.4 High-fat diet induces dyslipidemia, hepatic lipid accumulation and related protein expression changes in MAFLD mice. A: Changes in mouse body weight. B-E: Serum levels of TC, TG, LDL-C and FFA. F: Hepatic protein expression of CEBPA, SREBP-1c, FASN and ACC detected by Western blotting. G: Hepatic lipid accumulation observed by Oil Red O staining. The right panels show enlarged views (×400; Scale bar=50 μm) of the red dashed boxes in the left panels (×100; Scale bars=100 μm). Data are presented as Mean±SD (n=6). *P<0.05, **P<0.01, ***P<0.001 vs control group.

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