Search Results (422)

Metabolic dysfunction-associated steatohepatitis (MASH) represents a systems-level disorder driven by the interplay of metabolic stress, bile acid dysregulation, gut microbiota remodeling, and immune activation. Because no single experimental platform recapitulates the full spectrum of human disease—from steatosis and fibrosis to spontaneous hepatocellular carcinoma (HCC)—model selection must be guided by the dominant biological mechanism under investigation rather than by phenotypic similarity alone. This review proposes a mechanism-oriented framework for model selection, illustrated by representative experimental systems, including the intensified high-fat/high-cholesterol diet supplemented with cholate (iHFC diet) and Tsumura–Suzuki obese diabetic (TSOD) and non-obese (TSNO) mouse models. The iHFC diet provides a reproducible platform for interrogating the bile acid–microbiota–macrophage axis in fibro-inflammatory progression, whereas TSOD mice represent a valuable system in which spontaneous MASH–HCC development can emerge under chronic metabolic imbalance without engineered oncogenic triggers. TSNO mice serve as a controlled background for dissecting bile acid-dependent susceptibility. We further integrate hepatocyte mitochondrial dysfunction, immune remodeling, and stellate cell activation into this triadic framework and position additional diet-induced, genetic, and in vitro models within a complementary translational landscape. Together, this mechanism-centered framework provides a practical roadmap for rational model selection and enhanced translational precision in MASH and metabolic hepatocarcinogenesis research. Full article

Background: Cultured primary mouse hepatocytes undergo drastic phenotypic and metabolic reprogramming, while the temporal rules and regulatory machinery of membrane phospholipid remodeling remain elusive. Methods: Relying on a 0–72 h time-series in vitro culture system, this study integrated multi-omics technologies to dissect the temporal dynamics of phospholipid remodeling in hepatocytes. Through phosphatidylethanolamine N-methyltransferase (PEMT) knockout, exogenous PEMT expression, and methionine deprivation, we examined the association of PEMT status and methionine availability with phospholipid remodeling. Results: In vitro cultivation reduces intracellular total phospholipids, phosphatidylcholine (PC) and phosphatidylethanolamine (PE) through three coordinated events: suppressed transcription of phospholipid synthetic genes hinders de novo synthesis, elevated lipid hydrolysis consumes cellular phospholipids, and extracellular phospholipids accumulate in the culture medium from 12 to 48 h. These jointly trigger ordered remodeling of PC/PE balance, acyl chain length and fatty acid unsaturation. PEMT knockout was associated with PE retention without worsening hepatocyte dedifferentiation, PEMT exogenous expression raises PC content and PC/PE ratio yet cannot rescue culture-dominated lipid structural shifts. Methionine depletion depleted cellular methionine, S-adenosylmethionine (SAM) and S-adenosyl-L-homocysteine (SAH), producing selected lipid changes that partially overlapped with lipid phenotypes of PEMT knockout. Conclusion: In short, culture duration was the dominant factor associated with the fundamental phospholipid remodeling trajectory, and PEMT status and methionine availability, were associated with selective differences in lipid composition. Full article

Non-alcoholic fatty liver disease (NAFLD) is a prevalent metabolic disorder that can progress to non-alcoholic steatohepatitis (NASH), in which lipotoxicity, oxidative stress, apoptosis, and impaired autophagy contribute to liver injury. 14-Deoxy-11,12-didehydroandrographolide (deAND), a bioactive diterpenoid from Andrographis paniculata, has shown anti-inflammatory and antioxidant activities, but its role in NASH-associated lipotoxicity remains unclear. This study investigated the protective effects and underlying mechanisms of deAND using palmitic acid (PA)-treated AML12 hepatocytes and a choline-deficient, L-amino acid-defined, high-fat-diet (CDAHFD)-induced mouse model of NASH. In AML12 cells, PA impaired autophagic flux and reduced the expression of the mitophagy-associated proteins PINK1 and Parkin and increased p62, LC3-II, reactive oxygen species production, and apoptotic signaling. deAND treatment restored autophagic flux and increased PINK1 and Parkin expression, enhanced antioxidant defense-related proteins, including HO-1, GCLM, and GPX2, and reduced oxidative stress and apoptosis. The protective effects of deAND were attenuated by autophagy inhibitors, supporting the involvement of autophagy regulation. In CDAHFD-fed mice, deAND reduced hepatic steatosis, inflammation, fibrosis, apoptosis, and autophagy dysregulation. These findings suggest that deAND alleviates lipotoxic liver injury by restoring autophagic homeostasis and reducing oxidative stress. Full article

Deoxynivalenol (DON) and patulin (PAT) are common mycotoxins in cereals and fruits, posing health risks for animals and human beings. In order to study their liver toxicity, 24 mice were randomly assigned to four groups, with six replicates in each group (one mouse per cage). The mice were intragastrically administered with DON, PAT, DON + PAT (DP), or without DON and PAT (the control group) for 28 days, respectively. The results showed that body weight gain was significantly reduced by all toxin treatments, compared with the control group, and the lowest body weight gain was observed in the DP group. Histopathology revealed that hepatocyte damage and inflammatory infiltration were more serious in the DP group, exhibiting the highest mRNA abundances of MyD88, IFN-γ, and JAK2. The severity of hepatocyte apoptosis induced in each group followed the order: DON > DP > PAT; the severity of oxidative stress was ranked as DP > DON > PAT. Transcriptomic analysis revealed that numerous differentially expressed genes were regulated by DP treatment, which were mainly enriched in the MAPK, JAK-STAT, and transforming growth factor (TGF)-β signaling pathways. In conclusion, individual exposure to DON or PAT triggered hepatic injury, and their co-exposure further exacerbated liver damage. Full article

Objective: While Sanzi Sijun Formula (SSF) has exhibited preliminary efficacy against metabolic dysfunction-associated steatotic liver disease (MASLD), its mode of action remains undefined. This study therefore aimed to unravel its core therapeutic mechanisms. Methods: UPLC-MS was employed to characterize the major components of SSF. Male C57BL/6J mice were fed a high-fat diet combined with high-fructose/glucose drinking water (HFD-HF/G) for 10 weeks to establish a MASLD model, followed by SSF intervention. After 8-week treatment, body and liver weight, hepatic histopathological alterations, serum levels of lipids, transaminase, and inflammatory cytokines were detected, and transcriptomic sequencing was performed on mouse liver tissues for mechanistic exploration. AML12 hepatocytes stimulated with palmitic acid (PA) were treated with SSF alone or in combination with AMPK or SIRT1 specific inhibitors. RT-qPCR and Western blotting were used to detect the expression or activation levels of AMPK, SIRT1, and key lipid metabolism-related molecules. Results: A total of 77 active components were identified in SSF by UPLC-MS analysis. In MASLD model mice, SSF significantly reduced body and liver weight, serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-c), and alanine aminotransferase (ALT), suppressed the pro-inflammatory cytokines including TNF-α and IL-6, and elevated adiponectin levels. Histopathological staining demonstrated that SSF effectively alleviated hepatic steatosis, ballooning, and inflammatory cell infiltration. Transcriptomic profiling analysis verified the major regulatory effect of SSF on lipid metabolism and identified the AMPK/SIRT1 signaling pathway as a potential mechanism. Further experiments confirmed that SSF restored the levels of AMPK/ACC phosphorylation and SIRT1 expression, thereby modulating downstream lipid metabolism-related genes in liver tissues. In PA-induced AML12 cells, SSF significantly reduced intracellular accumulation of lipid and reactive oxygen species (ROS), which were partially abrogated by the inhibitors of AMPK or SIRT1. Conclusions: SSF exerts prominent effects against MASLD in both in vivo and in vitro models. Modulation of the AMPK/SIRT1 signaling pathway primarily contributes to its therapeutic mechanism against lipid metabolism disorder and lipotoxic liver injury. These findings provide experimental evidence to support the clinical application of SSF for MASLD treatment. Full article

An abnormality of Ca²⁺ signaling may aggravate lipid accumulation in steatotic hepatocytes, leading to non-alcoholic fatty liver disease. However, the molecular identity of Ca²⁺-permeable channels and the mechanism of involvement of these channels in steatotic hepatocytes are not well-studied. In the present study, we investigated the role of a Ca²⁺-permeable channel TRPM2 in lipid metabolism in steatotic hepatocytes. A mouse model of non-alcoholic fatty liver disease was established by high-fat-diet feeding. Fat accumulation, fibrosis, lipophagic indexes, TFEB and lysosomal acid lipase in the liver tissue and/or hepatocytes were compared between TRPM2-knockout mice and wild-type mice. Knockout of the TRPM2 gene aggravated liver fat accumulation and fibrosis. Mechanistically, the TRPM2 knockout impaired the lipophagic process, decreased lysosomal abundance and attenuated lysosomal/autolysosomal acidification in mouse hepatocytes. Furthermore, the TRPM2 knockout reduced TFEB expression and its nuclear translation and also reduced the expression/activity of lysosomal acid lipase. These data demonstrate that TRPM2 deficiency may reduce lipophagy via its action on TFEB and lysosomal acid lipase, consequently contributing to liver steatosis and NAFLD under high-fat feeding conditions. Full article

Background: Metabolic dysfunction-associated steatohepatitis (MASH), the progressive stage of metabolic dysfunction-associated steatotic liver disease (MASLD), is characterized by hepatic steatosis, inflammation, and fibrosis, yet no specific therapy has been established. This study evaluated the therapeutic potential of Andrias davidianus liver-derived peptides (ALPs) in a mouse model of MASH. Methods: ALPs were prepared by enzymatic hydrolysis of fresh Andrias davidianus liver. A MASH model was induced in mice using a methionine- and choline-deficient diet (MRCD). ALP was administered via oral gavage, and its effects were assessed through histological staining (H&E, Oil Red O, and Sirius Red), immunofluorescence (Ki67), apoptosis detection (TUNEL), serum biochemistry, and RNA-sequencing of liver tissues. Gut microbiota composition was also analyzed. Results: ALP treatment significantly alleviated hepatic histopathological features, including steatosis, inflammation, and fibrosis. It reduced aberrant proliferation and apoptosis of hepatocyte-like cells, and markedly improved serum biochemical markers of liver function. RNA-seq analysis revealed that ALP modulated the expression of lipid metabolism-related genes, an effect associated with suppression of the PPARγ signaling pathway. Furthermore, ALP selectively modulated MRCD-induced gut microbiota dysbiosis, particularly by reducing the Firmicutes-to-Bacteroidota (F/B) ratio and enriching Akkermansia. No overt toxicity was observed in other organs. Conclusions: Our findings demonstrate that ALP exerts protective effects against MASH by improving lipid metabolism, partially through suppression of the PPARγ signaling pathway, and by selectively modulating specific gut microbial taxa. ALP represents a promising natural therapeutic candidate for MASH. Full article

Background: Endotoxemia is a severe inflammatory condition that is characterized by acute immune responses and oxidative stress; endotoxemia can further develop into a cytokine storm and sepsis leading to severe organ damage. Our recent studies revealed that the growth hormone secretagogue receptor (GHSR) regulates macrophage polarization in obesity- and aging-associated chronic inflammation. However, its role in acute inflammation during endotoxemia remains unclear. Methods: We subjected myeloid-specific Ghsr knockout mice (LysM-Cre;Ghsr^f/f) to lipopolysaccharide (LPS)-induced endotoxemia in vivo and treated bone marrow-derived macrophages (BMDMs) with LPS in vitro. Subsequently, mouse survival rate and inflammatory signatures in the blood, peritoneal cavity, liver, and BMDM were assessed. In the ex vivo study, conditioned medium (CM) from BMDMs was applied to primary hepatocytes to assess how BMDM-derived CM influences hepatocyte inflammatory responses. Results: Myeloid-specific Ghsr knockout mice exhibited a significantly improved survival rate following LPS-induced endotoxemia, accompanied by reduced systemic inflammation, evident in the blood, peritoneal macrophages, and liver. In addition, Ghsr deficiency suppressed LPS-induced caspase-1 activation and pro-inflammatory cytokine secretion in macrophages. Consistent with these results, conditioned media from Ghsr-deficient BMDMs attenuated the inflammatory responses of primary hepatocytes. Mechanistically, LPS increased GHSR expression in BMDMs, and Ghsr-deficient BMDMs activated mitochondrial respiration and suppressed production of mitochondrial reactive oxygen species (ROS), resulting in downregulation of inflammatory activation of macrophages following LPS exposure. Conclusions: These data demonstrate that macrophage GHSR promotes systemic and tissue inflammation during endotoxemia by regulating mitochondria-associated macrophage polarization. The findings suggest that macrophage GHSR may represent a promising immunomodulatory target for acute inflammatory states, including endotoxemia and sepsis. Full article

Metabolic dysfunction-associated steatohepatitis (MASH) is a severe, progressive liver disease lacking effective therapies. Disulfiram (DSF), an FDA-approved medication for alcohol dependence, exhibits diverse biological activities beyond its primary indication. This study aimed to evaluate whether DSF holds intervention promise for MASH and to unravel the underlying molecular mechanism. The efficacy of DSF was assessed in a mouse model of MASH induced by a choline-deficient, L-amino acid-defined diet, as well as in hepatocytes exposed to free fatty acids (FFAs) to trigger lipotoxicity. RNA-seq analysis combined with bioinformatic approaches was performed to identify key pathways and hub genes. Mechanistic validation was carried out using Western blotting and qPCR. Computational predictions suggested that DSF may influence insulin resistance, inflammation, autophagy-related markers, and lipid metabolism. In FFAs-treated hepatocytes, DSF administration dose-dependently reduced lipid accumulation and lipotoxicity. Consistently, in MASH mice, DSF administration significantly lowered elevated serum ALT (35%) and AST (40%) levels and the absolute hepatic triglyceride content (reduced from 1 to 0.5 μg/mg protein), and markedly attenuated hepatic steatosis, inflammation, fibrosis, and oxidative stress. Of note, RNA-seq analysis revealed that DSF modulated autophagy-related pathways and identified Aurora kinase A (AURKA) as a central downregulated hub gene. Mechanistically, DSF suppressed AURKA expression, which in turn led to changes in autophagy-related markers. These changes in autophagy-related markers were functionally coupled to a reduction in lipotoxicity. Collectively, DSF alleviates MASH by inhibiting AURKA, thereby relieving AURKA-mediated suppression of autophagy-related markers, which was associated with diminishing lipotoxicity, and ultimately achieving broad suppression of disease progression. Thus, DSF represents a promising hepatoprotective candidate for the intervention of MASH. Full article

Background: Metabolic dysfunction-associated steatotic liver disease (MASLD) is now the most prevalent chronic liver disease worldwide, imposing a severe public health burden. Its core pathological hallmark is excessive hepatic lipid accumulation driven by systemic metabolic dysregulation. Concomitant hepatocellular injury impairs hepatic lactate clearance, leading to aberrant lactate buildup in the liver microenvironment. However, the causal role of lactate in exacerbating liver lipid metabolism dysfunction and driving the progression of MASLD remains unclear. Methods: First, we performed a comprehensive bioinformatic analysis of publicly available transcriptomic datasets. Mining of the Gene Expression Omnibus (GEO) database showed that lactate dehydrogenase (LDH) expression was significantly upregulated in liver tissues from both metabolic dysfunction-associated fatty liver disease (MASLD) patients and MASLD mouse models. Next, network pharmacology approaches were employed to predict putative molecular targets that could mediate lactate’s biological effects. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that these candidate targets were predominantly enriched in pathways governing fatty acid metabolism and long-chain fatty acid transport. Molecular docking and molecular dynamics simulations further suggested possible interactions and supported the prioritization of cluster of differentiation 36 (CD36) as candidate lipid metabolism regulators potentially involved in lactate-mediated effects. Finally, liver-specific Ldha knockdown mice (AAV8-TBG-shRNA) and free fatty acid-induced steatotic AML12 hepatocytes were used to investigate the functional relevance of these findings in vivo and in vitro. Results: Network pharmacology analyses preliminarily identified the PPAR signaling pathway as a candidate pathway potentially linking lactate to MASLD. Experimental results showed that exogenous lactate administration was associated with significantly increased lipid accumulation in steatotic AML12 hepatocytes and the livers of MASLD mice, manifested as elevated triglyceride levels and enhanced lipid droplet formation, accompanied by upregulated expression of PPARγ and CD36. Conversely, inhibiting endogenous lactate production or silencing PPARγ or CD36 attenuated this lipid-accumulation phenotype and significantly reduced intracellular triglyceride levels. Conclusions: In conclusion, these findings indicate that lactate exposure is associated with hepatic lipid accumulation and upregulation of the PPARγ/CD36 axis. Pharmacological inhibition or silencing of PPARγ or CD36 attenuates this phenotype, suggesting that this pathway may contribute to lactate-associated hepatic steatosis and potentially accelerate MASLD progression. Full article

The intestine, particularly the gut microbiota, and the liver play key roles in digestion, nutrient transformation, and immune regulation in fish. However, limited information is available regarding how different ornamental fish species regulate these systems under identical aquaculture conditions. Therefore, this study systematically compared gut microbiota diversity, structural variation, and predicted ecological functions among eight ornamental fish species reared in the same environment, using 16S rRNA high-throughput sequencing combined with digestive enzyme indices and histological analysis of intestinal and liver tissues. The results showed that goldfish (Carassius auratus) and crucian carp exhibited efficient digestive and absorptive capacities, supported by a thickened muscularis and prominent mucosal layers (p < 0.001). High goblet cell density was observed in red swordtail (Xiphophorus hellerii) and Mickey Mouse platy (Xiphophorus hellerii × X. maculatus) (p < 0.001). Larger hepatocyte perimeter and area were observed in red swordtail (p = 0.022, p = 0.015), whereas platinum mini parrot cichlid and sapphire mini parrot cichlid showed significant hepatocyte vacuolization. Microbial analysis showed that the eight fish species had similar α diversity indices, while the gut microbial profiles of Mickey Mouse platy and golden crucian carp differed the most. At the genus level, beneficial taxa such as Lactococcus, Paracoccus, and Cetobacterium were significantly enriched in red swordtail, sailfin molly, and goldfish, respectively, whereas opportunistic pathogens, including Edwardsiella, Aeromonas, and Acinetobacter, were enriched in Mickey Mouse platy, sapphire mini parrot cichlid, and golden crucian carp, respectively (p < 0.05). Functional prediction based on KEGG pathways indicated that sailfin molly and Mickey Mouse platy exhibited the broadest functional enrichment, primarily involving amino acid metabolism, fatty acid metabolism, and antibiotic biosynthesis. Crucian carp and golden crucian carp showed higher activity in amino acid biosynthesis and glycolysis/gluconeogenesis pathways. The two parrot cichlid species were characterized by enrichment in biofilm formation pathways of pathogenic bacteria and amino sugar and nucleotide sugar metabolism pathways. Goldfish and red swordtail were mainly associated with quorum sensing and ABC transporter pathways. These results provide a theoretical foundation for optimizing aquaculture conditions for ornamental fish and improving fish health and production efficiency. Full article

Tobacco leaf extract (TLE) exposure can induce liver injury-associated responses involving cell death, inflammatory signaling, and extracellular matrix (ECM)-related changes. This study examined whether apple seed extract (ASE) post-treatment changes apoptosis-, inflammation-, ECM-, and insulin-like growth factor (IGF)-related markers after TLE exposure. Primary mouse hepatocytes were exposed to TLE, ASE alone, or TLE followed by ASE, and mouse liver tissues were examined after TLE exposure with or without ASE post-treatment. TLE reduced hepatocyte viability in a concentration-dependent manner, with an IC₅₀ of 4.4 mg/mL. Annexin V/propidium iodide analysis showed that early apoptosis increased from 2.20% in untreated cells to 5.50% after 2 mg/mL TLE and 85.65% after 4 mg/mL TLE. ASE alone at 40 µg/mL increased the early apoptotic fraction to 53.65%, indicating that this concentration was not biologically neutral under basal culture conditions. After TLE exposure followed by ASE post-treatment, the live-cell fraction remained high in T2HA and T4HA, whereas T5HA retained a high early apoptotic fraction. In mice, TLE exposure was accompanied by visible liver appearance changes and histological alterations. ASE post-treatment changed Alcian blue staining, gelatinase activity, TIMP-associated signals, and IGF-related signals. These findings indicate treatment-dependent changes in selected injury-associated markers rather than a consistently protective effect of ASE. The study does not assign these effects to a specific ASE constituent because compound-level chemical standardization was not performed. Full article

Background: Acetaminophen (APAP) overdose remains a major cause of acute liver injury. Although N-acetylcysteine (NAC) is the clinically established antidote for APAP toxicity, its efficacy is greatest when administered early, and additional therapeutic strategies are still needed for patients with delayed presentation or progressive injury. Because APAP hepatotoxicity involves coupled disturbances in redox control, mitochondrial performance, and cellular metabolism, metabolic enzymes that sustain NADPH availability may critically influence disease severity. Malic enzyme 1 (ME1), a cytosolic NADPH-generating enzyme, has not been functionally defined in this context. Methods: To determine the contribution of ME1 to APAP-induced liver injury (AILI), we used hepatocyte-specific ME1 knockout mice, hepatic overexpression and reconstitution approaches, primary mouse hepatocytes, and an enzymatically inactive ME1 mutant. Liver injury and associated changes in oxidative stress, mitochondrial function, energy metabolism, autophagic flux, and endoplasmic reticulum (ER) stress were evaluated using biochemical, histological, molecular, and ultrastructural analyses, together with pharmacological interventions. Results: Genetic loss of ME1 did not substantially alter early APAP metabolic activation-related indices, including APAP-protein adduct formation, but markedly increased hepatocellular metabolic vulnerability after APAP challenge. This phenotype was characterized by enhanced lipid peroxidation, impaired mitochondrial polarization, reduced ATP availability, defective autophagic flux, and amplified ER stress, leading to more severe liver damage. In contrast, ME1 overexpression or reconstitution promoted a more adaptive metabolic response and limited tissue injury. These effects depended largely on ME1 catalytic activity, as protection was markedly weakened with the mutant enzyme. Pharmacological analyses further supported the involvement of AMPK/mTOR-associated autophagy regulation and ER stress adaptation in the downstream actions of ME1. Malic acid also partially attenuated APAP-induced hepatotoxicity in vivo and in vitro. Conclusions: ME1 functions as an endogenous metabolic factor that influences the outcome of APAP-induced liver injury. Its catalytic activity supports hepatocyte survival primarily by preserving reductive capacity, bioenergetic balance, and adaptive stress responses, rather than by altering APAP metabolic activation. Full article

Metabolic dysfunction-associated steatotic liver disease (MASLD) is highly associated with the development of cardiovascular disease (CVD); however, the mechanisms responsible are currently unknown. We have developed a model of MASLD due to the loss of hepatocyte peroxisome proliferator-activated receptor α (PPARα^HEPKO). We found that plasma beta-hydroxybutyrate (BHOB) levels were significantly reduced in PPARα^HEPKO mice and aimed to investigate the therapeutic potential of restoring BHOB levels in the development of CVD in these mice. Thirty-week-old PPARα^HEPKO and control PPARα^FL/FL mice were randomized to receive 1,3 butanediol (1,3-BDO), a precursor of BHOB, in drinking water for 6 weeks. 1,3-BDO treatment resulted in a significant increase in plasma BHOB levels, a significant decrease in mean arterial blood pressure, improvement in systolic and diastolic function, a decrease in vascular stiffness, and improved exercise performance in PPARα^HEPKO mice. 1,3-BDO treatment did not alleviate hepatic steatosis in PPARα^HEPKO mice; however, it improved plasma cholesterol levels and decreased cardiac lipid accumulation, fibrosis, and apoptosis. 1,3-BDO treatment also resulted in a significant increase in cardiac AMP-activated protein kinase (AMPK) levels. Increasing plasma BHOB levels reverses CVD in our mouse model of MASLD. A similar approach could be an effective strategy for preventing the development of CVD in patients with human MASLD. Full article

Non-coding RNAs are pivotal regulators of metabolic disease pathogenesis, yet the role of microRNA-27a (miR-27a) in obesity-associated hepatic steatosis remains incompletely characterized. This study examined the functional contribution and molecular mechanism of miR-27a in regulating hepatocyte mitochondrial homeostasis and lipid metabolism. Utilizing in vivo mouse models, including low-fat diet controls, high-fat diet (HFD)-induced obesity, and gain- and loss-of-function approaches, miR-27a was found to be markedly upregulated in the serum and liver of obese mice, correlating with disrupted glucose and lipid homeostasis as well as hepatic steatosis. Mechanistically, miR-27a overexpression recapitulated HFD-induced mitochondrial dysfunction, manifested by decreased mitochondrial biogenesis and elevated reactive oxygen species (ROS) production. Conversely, genetic silencing of miR-27a restored mitochondrial integrity and mitigated lipid accumulation. In vitro experiments using HepG2 cells confirmed that miR-27a directly suppresses nuclear factor erythroid 2-related factor 2 (NFE2L2), and NFE2L2 overexpression counteracted miR-27a-induced mitochondrial damage and steatosis. Collectively, these results demonstrate that miR-27a promotes hepatic steatosis by targeting NFE2L2, leading to mitochondrial impairment and oxidative stress, highlighting miR-27a as a potential biomarker and therapeutic target for obesity-associated liver metabolic disorders. Full article