Systemic lipid remodeling associated with deferoxamine treatment, dietary intervention and combinatorial effect in ameliorating high cholesterol diet-induced steatotic liver in mice.

Abstract

INTRODUCTION: Metabolic dysfunction-associated steatotic liver disease (MASLD) is becoming increasingly more prevalent and poses a major growing health concern worldwide. Iron deposition have been observed in the liver of MASLD patients and iron can lead to the formation of reactive oxygen species. Although lipids were altered in the livers of MASLD patients, the underlying mechanisms implicated in this disease have not yet been fully elucidated. OBJECTIVES: To investigate lipid alterations in MASLD during deferoxamine (DFO, an iron chelator) and/or dietary interventions and to unravel the underpinning pathways involved in this disease. METHODS: In this study, C57BL/6N mice were subjected to high cholesterol diet (HCD) to induce steatotic liver seen in MASLD. This study systematically examined the effects of DFO treatment, dietary intervention, and their combinatorial impacts on steatotic liver phenotype, lipidome and proteome. We performed hematoxylin & eosin staining, alanine amino transferase activity assay, oleic acid and cholesterol uptake assay, gene siliencing, filipin staining, iron assay, inflammatory marker measurements and utilized multi-omics technologies (lipidomics and proteomics using mass spectrometers) to interrogate MASLD. RESULTS: Comprehensive lipidomics analysis revealed that elevated GM3 levels resulting in abnormal lipid metabolism in the HCD-fed mice decreased after DFO treatment and dietary intervention. Proteomics analysis uncovered significant decreases in the expression levels of hepatic proteins involved in both biosynthesis and uptake of fatty acid and cholesterol after DFO treatment. These results were verified with isotope labeled oleic acid and/or cholesterol uptake assay. Furthermore, the iron chelating properties of DFO alleviated oxidative stress induced by HCD. The pathways of "Cytoplasmic ribosomal proteins"and "G13 signaling" were restored after DFO treatment and dietary intervention. CONCLUSION: The combinatorial synergistic effect of diet and DFO successfully restored LPC and GM3 levels back to almost control levels in MASLD mice as well as restored lipid and protein homeostasis. Ultimately, culminating in the alleviation of MASLD triggered dysregulated pathway.