High dietary Fructose Drives Metabolic Dysfunction-Associated Steatotic Liver Disease via Activating ubiquitin-specific peptidase 2/11β-hydroxysteroid dehydrogenase type 1 Pathway in Mice - PubMed (original) (raw)
High dietary Fructose Drives Metabolic Dysfunction-Associated Steatotic Liver Disease via Activating ubiquitin-specific peptidase 2/11β-hydroxysteroid dehydrogenase type 1 Pathway in Mice
Chunlin Li et al. Int J Biol Sci. 2024.
Abstract
Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most common cause of chronic liver-related morbidity and mortality. Though high fructose intake is acknowledged as a metabolic hazard, its role in the etiology of MASLD requires further clarification. Here, we demonstrated that high dietary fructose drives MASLD development and promotes MASLD progression in mice, and identified Usp2 as a fructose-responsive gene in the liver. Elevated USP2 levels were detected in the hepatocytes of MASLD mice; a similar increase was observed following fructose exposure in primary hepatocytes and mouse AML12 cells. Notably, hepatocytes overexpressing USP2 presented with exaggerated lipid accumulation and metabolic inflammation when exposed to fructose. Conversely, USP2 knockdown mitigated these fructose-induced changes. Furthermore, USP2 was found to activate the C/EBPα/11β-HSD1 signaling, which further impacted the equilibrium of cortisol and cortisone in the circulation of mice. Collectively, our findings revealed the role of dietary fructose in MASLD pathogenesis and identified the USP2-mediated C/EBPα/ 11β-HSD1 signaling as a potential target for the management of MASLD.
Keywords: MASLD; USP2; dietary fructose; hepatic steatosis; inflammation.
© The author(s).
Conflict of interest statement
Competing Interests: The authors have declared that no competing interest exists.
Figures
Figure 1
High dietary fructose drives the development of MASLD (A) The design of animal experiment; (B) Dynamic changes of body weight of mice; (C) Liver-to-body weight ratio; (D) Pathological staining of liver (magnification 400×): H&E staining and ORO staining of liver sections (magnification 400×); (E) NAS score; (F) Serum levels of TG, CHOL, and LDL-c; (G) Serum levels of ALT, AST, and ALP. Data are presented as mean ± SD. Con vs WDF: *; Con vs Fr: #. *p<0.05, **p<0.01, ***p<0.001; #p<0.05, ##p<0.01, ###p<0.001.
Figure 2
High dietary fructose promotes the progression of MASLD (A) Fructose induction scheme with WD feeding; (B) Dynamic changes in body weight of mice; (C) Liver-to-body weight ratio; (D) Pathological staining of liver (magnification 400×): H&E staining and ORO staining of liver sections (magnification 400×); (E) NAS score; (F) Serum levels of TG, CHOL, and LDL-c; (G) Serum levels of ALT, AST, and ALP. The quantification data are presented as mean ± SD. Con vs WD: *; WD vs WFr: #. *p<0.05, **p<0.01, ***p<0.001; #p<0.05, ##p<0.01, ###p<0.001.
Figure 3
Fructose upregulates hepatic USP2 expression (A) Plot of PLSDA analysis between Con and Fr mice; (B) Heatmap of DEGs (p<0.05) between Con and Fr mice; (C) KEGG analysis between Con and Fr groups; (D) Plot of PLSDA analysis between WD and WDF groups. (E) Heatmap of DEGs (p<0.05) between WD and WDF groups; (F) KEGG analysis between WD and WDF groups; (G) Volcano plot of significant DEGs (p adj<0.05) of Fr vs Con, WDF vs WD, and WD vs Con, respectively; (H) Plot of co-responsive genes of Fr and WDF groups when compared to Con and WD groups, respectively; (I) Relative mRNA level of Usp2 gene in the liver; (J) Protein blotting of USP2 in the liver; (K) The immunohistochemical staining in the liver for USP2 protein (magnification 400×) and the quantification of the positively stained area. The quantification data are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001.
Figure 4
Fructose increases lipid accumulation and USP2 expression in primary hepatocytes. (A) The cell experiment flowchart; (B) The TG content of primary hepatocytes; (C) The Lipi-Red staining of primary hepatocytes (magnification 200×), the bottom panel figures are amplification of the upper panel; (D) The ORO staining of primary hepatocytes (magnification 200×), the bottom panel figures are amplification of the upper panel; (E) The levels of IL-1β, IL-6, and TNF-α in the culture medium of primary hepatocytes; (F) The immunofluorescence for USP2 in primary hepatocytes (magnification 100×); (G) Relative mRNA level of Usp2 gene in primary hepatocytes; (H) Protein blotting of USP2 in primary hepatocytes. The quantification data are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001.
Figure 5
Fructose induces hepatocyte steatosis and inflammation via USP2 in primary hepatocytes. (A) The cell experiment flowchart of Usp2 overexpression in primary hepatocytes; (B) The mRNA expression of Usp2 gene in primary hepatocytes; (C) The Lipi-Red staining (magnification 200×) of _Usp2_-overexpressed primary hepatocytes, the bottom panel figures are amplification of the upper panel; (D) The ORO staining (magnification 100×) of _Usp2_-overexpressed primary hepatocytes, the bottom panel figures are amplification of the upper panel; (E) The TG content of _Usp2_-overexpressed primary hepatocytes; (F) The levels of IL-1β, IL-6, and TNF-α in the culture medium of _Usp2_-overexpressed primary hepatocytes; (G) The cell experiment flowchart of Usp2 knockdown in primary hepatocytes; (H) The mRNA expression of Usp2 gene in primary hepatocytes; (I) The Lipi-Red staining (magnification 200×) of _Usp2-_knockdowned primary hepatocytes, the bottom panel figures are amplification of the upper panel; (J) The ORO staining (magnification 100×) of _Usp2-_knockdowned primary hepatocytes, the bottom panel figures are amplification of the upper panel; (K) The TG content of _Usp2-_knockdowned primary hepatocytes; (L) The levels of IL-1β, IL-6, and TNF-α in the culture medium of _Usp2-_knockdowned primary hepatocytes. The quantification data are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001.
Figure 6
The function of USP2 depends on C/EBPα/ 11β-HSD1 in fructose-stressed primary hepatocytes. (A) Hypothesis diagram for USP2/ 11β-HSD1 pathway in the liver; (B, C) Relative mRNA expression of C/EBPα (C/ebpα) and 11β-HSD1 (Hsd11b1) in hepatocytes; (D) Protein blotting of C/EBPα and 11β-HSD1 in hepatocytes; (E) Relative mRNA expression of USP2 (Usp2), C/EBPα (C/ebpα), and 11β-HSD1 (Hsd11b1) in _Usp2_-overexpressed hepatocytes; (F) Protein blotting of USP2, C/EBPα, and 11β-HSD1 in _Usp2_-overexpressed hepatocytes; (G) Relative mRNA expression of USP2 (Usp2), C/EBPα (C/ebpα), and 11β-HSD1 (Hsd11b1) in _Usp2_-knockdowded hepatocytes; (H) Protein blotting of USP2, C/EBPα, and 11β-HSD1 in _Usp2_-knockdowded hepatocytes. The quantification data are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001.
Figure 7
Fructose induces MASLD via the USP2/ 11β-HSD1 pathway (A, B) Relative mRNA expression of C/EBPα (C/ebpα) and 11β-HSD1 (Hsd11b1) in the liver of the mice; (C) Protein blotting and statistical analysis of C/EBPα and 11β-HSD1 in the liver of the mice; (D) Serum levels of cortisol and cortisone, and cortisone-to-cortisol ratio. The quantification data are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001.
References
- Rinella ME, Lazarus JV, Ratziu V, Francque SM, Sanyal AJ, Kanwal F. et al. A multisociety Delphi consensus statement on new fatty liver disease nomenclature. J Hepatol. 2023;79:1542–56. - PubMed
- Lim GEH, Tang A, Ng CH, Chin YH, Lim WH, Tan DJH. et al. An Observational Data Meta-analysis on the Differences in Prevalence and Risk Factors Between MAFLD vs NAFLD. Clin Gastroenterol Hepatol. 2023;21:619–29.e7. - PubMed
- Younossi ZM, Zelber-Sagi S, Henry L, Gerber LH. Lifestyle interventions in nonalcoholic fatty liver disease. Nat Rev Gastroenterol Hepatol. 2023;20:708–22. - PubMed
- Lê KA, Ith M, Kreis R, Faeh D, Bortolotti M, Tran C. et al. Fructose overconsumption causes dyslipidemia and ectopic lipid deposition in healthy subjects with and without a family history of type 2 diabetes. Am J Clin Nutr. 2009;89:1760–5. - PubMed
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