Search Results (4,647)

Metabolic dysfunction-associated steatotic liver disease (MASLD) and its progressive form, metabolic dysfunction-associated steatohepatitis (MASH), represent a growing global health burden driven by complex interactions among hepatic lipid accumulation, insulin resistance, chronic inflammation, and mitochondrial dysfunction. Exercise remains the cornerstone of lifestyle therapy for MASLD/MASH; however, its therapeutic benefits extend well beyond weight reduction and involve coordinated molecular adaptations across multiple organ systems. In this review, we introduce hepatic mitochondrial reprogramming as a conceptual framework describing the coordinated remodeling of mitochondrial energetics, quality-control pathways, and redox homeostasis that collectively restore metabolic flexibility and hepatocellular resilience. Exercise activates key metabolic regulators, including AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), and sirtuin signaling, promoting mitochondrial biogenesis, fatty acid oxidation, oxidative phosphorylation, and mitophagy while suppressing hepatic lipogenesis and oxidative injury. Skeletal muscle-derived myokines, alterations in gut microbial metabolism, and thyroid hormone signaling converge upon hepatic mitochondrial function through complementary endocrine and metabolic pathways. Together, these adaptations reduce hepatic steatosis, lipotoxicity, inflammation, and fibrogenesis while improving insulin sensitivity and metabolic flexibility. Emerging evidence further suggests that exercise-induced mitochondrial remodeling may complement pharmacologic therapies targeting hepatic metabolism, including thyroid hormone receptor-β agonists. Although multi-omics technologies continue to expand our understanding of these adaptive responses, the present review emphasizes the underlying molecular and physiological mechanisms through which exercise remodels hepatic mitochondrial function. We propose that exercise acts as a systems-level mitochondrial remodeling stimulus integrating skeletal muscle-, gut-, and thyroid-derived signals to improve hepatic metabolism and attenuate MASLD/MASH progression. This conceptual framework provides a mechanistic basis for precision exercise prescriptions and future combination therapeutic strategies targeting mitochondrial health. Full article

Objective: The objective of this study is to investigate the clinical value of the triglyceride-glucose (TyG) index combined with alanine aminotransferase (ALT) in identifying prevalent metabolic-associated fatty liver disease (MAFLD) in patients with obstructive sleep apnea (OSA), and to construct and validate a diagnostic risk-stratification model. Methods: Clinical data of OSA patients were retrospectively collected from two centers: the Department of Geriatrics, Peking University People’s Hospital (August 2021 to December 2025) and the Department of Geriatrics, Shijiazhuang People’s Hospital (June 2023 to December 2025). MAFLD was diagnosed by abdominal ultrasonography performed by experienced radiologists blinded to laboratory results. Candidate predictors were selected using univariate logistic regression, LASSO regression, and bootstrap stability testing. Model performance was assessed by discrimination (area under the receiver operating characteristic curve, AUC), calibration (Hosmer–Lemeshow test, calibration curves), and clinical utility (decision curve analysis). Internal validation was performed using 10-fold cross-validation and bootstrap resampling with optimism correction. External validation was conducted in an independent cohort. Sensitivity analyses included subgroup analyses stratified by sex, age, BMI, and OSA severity. Results: A total of 962 patients were included in the development set and 116 in the external validation set. Multivariate analysis identified TyG index (OR = 1.95, 95% CI: 1.51–2.53), LDL-C (OR = 1.24, 95% CI: 1.02–1.49), BMI (OR = 1.21, 95% CI: 1.16–1.26), and ALT (OR = 1.04, 95% CI: 1.02–1.05) as variables independently associated with prevalent MAFLD, while platelet-to-lymphocyte ratio (PLR) was protective (OR = 0.996, 95% CI: 0.993–0.999). The simplified TyG-ALT model achieved an AUC of 0.714 (95% CI: 0.685–0.744) in the development set, with an optimism-corrected AUC of 0.712, and an AUC of 0.783 (95% CI: 0.701–0.866) in external validation. The model demonstrated good calibration and favorable clinical net benefit within the threshold range of 0.30–0.70. The optimal cutoff was 0.564, with sensitivity of 68.7% and specificity of 79.6%. Conclusions: The TyG-ALT model demonstrates good discriminative ability, calibration, and clinical utility for case-finding and risk stratification of prevalent MAFLD in OSA patients, particularly for identifying high-risk individuals requiring confirmatory imaging. Full article

Background and Objective: Non-alcoholic fatty liver disease (NAFLD) induces systemic low-grade inflammation and lipotoxicity. While its microvascular effects on the inner ear are recognized, its impact on the middle ear remains unexplored. This study aimed to investigate subclinical biomechanical alterations in the middle ear of normal-hearing NAFLD patients using Wideband Tympanometry (WBT). Materials and Methods: This prospective observational study included 28 patients with ultrasonographically confirmed NAFLD and 30 healthy, sex-matched controls. All included participants had normal pure-tone hearing thresholds and Type A tympanograms. WBT was performed to measure Resonance Frequency (RF) and frequency-specific absorbance (226–8000 Hz). A subgroup analysis compared early-stage (Grade 1) and advanced-stage (Grades 2–3) hepatosteatosis. Results: The mean RF was significantly lower in the NAFLD group compared to the controls for both the right (781.6 ± 218.7 vs. 1010.6 ± 529.0 Hz; p = 0.035) and left ears (740.3 ± 212.7 vs. 1221.6 ± 754.2 Hz; p = 0.002). Furthermore, wideband absorbance was significantly elevated at lower frequencies (250 Hz and 500 Hz) in the study group (p < 0.001 for both ears). Middle ear resting pressures were similar between the groups. Subgroup analysis revealed no significant differences in WBT parameters between Grade 1 and advanced stages (p > 0.05). Conclusions: Normal-hearing NAFLD patients exhibit a mass-dominated biomechanical shift in the middle ear, characterized by a decreased RF and increased low-frequency absorbance. These subclinical alterations, observable even in early-stage hepatosteatosis, likely reflect mucosal micro-edema driven by systemic inflammation. WBT serves as a sensitive, non-invasive tool for detecting early metabolic-related otologic changes. Full article

Introduction: Hepatic fibrosis represents a critical intermediate stage in the progression from chronic liver disease to cirrhosis and ultimately hepatocellular carcinoma. Spirulina platensis, a microorganism rich in bioactive compounds, contains several functional components, among which are Spirulina polysaccharides and phycocyanin. Both have been demonstrated to exhibit multiple biological activities, including antioxidant, anti-inflammatory, and immunomodulatory effects. However, the intervention effects and mechanisms of Spirulina polysaccharides and phycocyanin on CCl₄-induced hepatic fibrosis remain unclear. 16S rRNA sequencing and non-targeted metabolomics technologies are employed to analyze bacterial taxa and metabolic pathways. Methods: Hepatic fibrosis (HF) was induced in male C57BL/6J mice via intraperitoneal injection of CCl₄. After 1 week of modeling, mice were intragastrically administered SPP or PC for 4 consecutive weeks. Gut microbiota composition and fecal metabolites were analyzed using 16S rRNA gene sequencing and metabolomics. Results: SPP showed more pronounced protective effects than PC under the experimental conditions used in this study. SPP treatment significantly alleviated hepatic fibrosis in mice. Compared with the model group, SPP administration markedly increased the abundance of bacterial taxa and modulated fecal metabolic profiles, including short-chain fatty acid metabolism. Conclusions: SPP treatment mitigates CCl₄-induced hepatic fibrosis in mice. These protective effects may be associated with the modulation of gut microbiota composition and fecal metabolic pathways, including those related to short-chain fatty acid metabolism. Full article

Background/Objectives: The glucocorticoid receptor (GR) is essential for regulating liver energy balance, metabolism, and inflammation. Stress and other factors can impair its function, resulting in glucocorticoid-resistant metabolic liver disease. GR mainly exists in two forms: the glucocorticoid-binding isoform, GRα, and the non-binding isoform, GRβ. The GRβ isoform typically exhibits minimal signaling activity beyond its role as a dominant-negative regulator of GRα, thereby decreasing glucocorticoid responsiveness and potentially causing resistance. Methods: To explore GRβ signaling independent of GRα, we developed mice with adenovirus-induced overexpression of GRβ (GRβ-Ad) and control mice with a vector (Vec-Ad). After five days on a standard diet, these mice received either vehicle or dexamethasone treatment. Liver tissues were collected, and we performed RNA sequencing and advanced PamGene kinome analysis to detect pathway changes in GRβ-Ad mice compared with controls. Results: Significant increases were observed in the expression of genes that inhibit fatty acid oxidation, inflammation, and liver cancer development. There was also a marked difference in serine/threonine kinase activity between GRβ-Ad and control mice. Conclusions: The findings suggest that elevated GRβ levels affect kinase pathways that modulate glucocorticoid signaling, disrupt liver lipid metabolism, and are associated with cancer pathways. Further research is needed to determine whether GRβ functions similarly in humans and to assess its potential contribution to hepatocellular carcinoma (HCC). Full article

Metabolic dysfunction-associated fatty liver disease (MAFLD) is now the leading cause of hepatocellular carcinoma (HCC) globally. HCC surveillance is currently restricted to patients with cirrhosis, leaving those without cirrhosis, who present with more advanced disease and poorer outcomes without adequate risk stratification tools. This study compared lipid profiles across MAFLD and MAFLD-related HCC (MAFLD-HCC) patients, with and without cirrhosis, to characterise metabolic dysregulation underlying non-cirrhotic MAFLD-HCC (ncMAFLD-HCC). Plasma and liver lipidomic profiles were obtained from 221 patients (140 MAFLD, 66 cirrhotic MAFLD-HCC (cMAFLD-HCC), and 15 ncMAFLD-HCC) using untargeted liquid chromatography mass spectrometry. Univariate, multivariable and enrichment analyses were performed for statistically determining the lipid profile difference between the groups. Seventy percent of lipid classes were more abundant in MAFLD than in ncMAFLD-HCC and cMAFLD-HCC. Multivariate analysis revealed distinct lipid profiles across the three groups in both plasma and liver. Over 100 lipid species including diglyceride (DAG), sphingomyelin (SM), triglyceride (TG), dihydroceramide (DHCer), and linoleic acid derivatives were differentially expressed in ncMAFLD-HCC versus MAFLD, with enrichment in pathways such as glycerolipid metabolism, G-protein signalling, MAPK signalling, EGFR-TKI resistance pathway, implicated in HCC development. ncMAFLD-HCC exhibits a distinct lipid signature, offering preliminary mechanistic insight and a foundation for non-invasive biomarker development. Full article

The rising burden of diabetic kidney disease (DKD) and its associated lipid abnormalities underscores the need for new mechanistic insights. Uromodulin, a kidney-enriched protein, has been associated with metabolic disorders in human studies, yet its functional role in systemic lipid metabolism remains elusive. In this study, transcriptomic datasets were analyzed to investigate uromodulin expression and biological function in DKD. Subsequently, a diabetic model was induced in UMOD^+/+ and UMOD^−/− rats using a combination of a high-fat diet, unilateral nephrectomy, and streptozotocin to assess renal and metabolic phenotypes. Public RNA-seq data indicated that uromodulin expression was downregulated in DKD and was enriched in the fatty acid metabolism pathway. At baseline, UMOD^−/− rats resembled UMOD^+/+ rats in terms of growth, routine serum lipids, and major organ function. However, in diabetes, UMOD^−/− rats exhibited higher mortality and pronounced hyperlipidemia. Hyperlipidemia occurred prior to the onset of renal dysfunction. Of note, this exacerbated lipid dysregulation represented a lipodystrophy-like phenotype rather than secondary changes in the pancreas, liver, or circulating cytokines (IL-6, IL-1β, and TNF-α). Moreover, UMOD^−/− rats displayed exacerbated tubular injury and enhanced renal lipid accumulation in DKD relative to UMOD^+/+ rats. Collectively, uromodulin protects diabetic rats from death, prevents epididymal white adipose tissue from browning, and attenuates kidney injury. Our findings identify uromodulin as a novel regulator of the kidney–adipose axis. Full article

Alcohol-related liver disease (ALD) poses a significant global health burden, driven by complex mechanisms including oxidative stress, inflammation, and gut–liver axis disruption. While the individual roles of gut microbiota dysbiosis and ageing in ALD pathogenesis are increasingly recognized, their synergistic interaction remains poorly understood. This review synthesizes current evidence to argue that there is an interaction between ageing and the gut microbiota that collectively amplifies progression of ALD. Specifically, ageing promotes gut dysbiosis through immunosenescence (e.g., reduced IgA diversification and antimicrobial peptide decline), intestinal barrier failure, and altered microbial metabolite profiles (e.g., decreased short-chain fatty acids and dysregulated bile acid metabolism). Conversely, dysbiosis-derived metabolites and endotoxins modulate ageing-related signaling pathways, including SIRT1, FOXO, and Nrf2, thereby accelerating hepatic cellular senescence, inflammation, and fibrogenesis. Furthermore, we also discussed the typical microbial changes in ALD. These include an increase in the Proteobacteria, a decrease in the Bacteroidetes, as well as imbalances in fungi and viruses. In ageing, similar but distinct shifts occur, such as reduced microbial diversity, decreased short-chain fatty acid producers, and increased intestinal permeability. Therapeutic strategies targeting the gut microbiota (probiotics, fecal microbiota transplantation) or ageing-related pathways (SIRT1 activators) hold promise. Future research priorities include validating ageing-associated microbial signatures as predictors of ALD progression and testing microbiota-targeted interventions in aged preclinical models. Collectively, this review identifies the microbiota–ageing axis as a tractable therapeutic target for ALD and provides a framework for future mechanistic and translational studies. Full article

Background: Type 2 diabetes mellitus (T2DM) is associated with subclinical myocardial dysfunction, which may occur despite preserved left ventricular ejection fraction. Left ventricular global longitudinal strain (LV GLS) is a sensitive marker of early systolic impairment and may detect subtle changes in myocardial function before conventional echocardiographic parameters become abnormal. The effect of sodium-glucose cotransporter-2 inhibitors (SGLT2i) on LV GLS in adults with T2DM remains incompletely defined. Objective: To synthesize the available evidence on the effects of SGLT2i therapy on LV GLS or LV strain in adults with T2DM. Methods: Original full-text human studies evaluating SGLT2i therapy in adults with T2DM and reporting LV GLS or LV strain were included. LV GLS was assessed primarily by speckle-tracking echocardiography, while one study used cardiac magnetic resonance feature-tracking. Reviews, conference abstracts, protocols, animal-only studies, and studies without LV strain assessment were excluded. Risk of bias was assessed using RoB 2 for randomized studies and ROBINS-I for non-randomized studies. Results: Twenty-six studies involving more than 2300 participants were included. The studies evaluated dapagliflozin, empagliflozin, ertugliflozin, canagliflozin, or mixed SGLT2i regimens across heterogeneous clinical populations, including patients with preserved ejection fraction, pre-heart failure, diabetes-related cardiomyopathy, chronic heart failure, coronary artery disease, hypertension, non-alcoholic fatty liver disease, and cardio-oncology risk. Most observational and before–after studies reported favorable changes in LV GLS after SGLT2i therapy, whereas randomized and controlled studies showed more variable findings. Several studies also reported improvements in LV remodeling, diastolic function, left atrial function, myocardial work indices, NT-proBNP, cardiometabolic parameters, or epicardial adipose tissue thickness. However, the certainty of evidence was limited by methodological heterogeneity, differences in comparator groups, variable follow-up duration, non-standardized imaging protocols, and risk of bias, particularly in non-randomized and single-arm studies. Conclusions: SGLT2i therapy may be associated with favorable changes in LV GLS in adults with T2DM, suggesting a potential beneficial effect on subclinical left ventricular systolic function. However, current evidence does not definitively establish a consistent treatment effect across all populations. Larger randomized controlled trials with standardized strain imaging protocols, predefined LV GLS endpoints, and clinically relevant follow-up are needed to determine whether SGLT2i-related improvements in LV GLS reflect true myocardial benefit and translate into improved cardiovascular outcomes. Full article

Background: Metabolic dysfunction-associated steatotic liver disease (MASLD) has been linked to atrial fibrillation (AF); however, its relationship with specific AF subtypes remains unclear. This prospective, single-center, observational case–control study investigated whether MASLD is independently associated with AF presence and its subtypes. Materials: A total of 327 participants were analyzed, including 119 controls and 208 patients with AF. Comprehensive clinical history, anthropometric measures, laboratory testing, 24 h Holter ECG, and echocardiography were performed. Clinically inferred MASLD was defined according to the current EASL–EASD–EASO guidelines using clinical and non-invasive indices (Hepatic Steatosis Index, Fatty Liver Index, Fibrosis-4 Index). No liver biopsy or imaging confirmation of steatosis or fibrosis was performed, and therefore, the diagnosis represents a clinically inferred (“probable”) MASLD. To minimize systematic bias and improve baseline comparability between groups, propensity score matching and complementary regression analyses were applied. Results: Overall probable MASLD prevalence did not differ between AF and controls (42% vs. 44%, p = 0.742). A clear phenotypic gradient emerged across subtypes: lowest in permanent AF (PermAF, 27.1%) versus paroxysmal (47.1%) and persistent AF (51.4%) (p = 0.021). PermAF exhibited the most advanced comorbidity—highest CHF (78.6%), CKD (71.4%), HFpEF (48.6%), FIB-4 (median 2.67), the lowest TG/HDL–cholesterol ratio (1.93 vs. 3.32; p < 0.001), and progressive renal impairment. Statin therapy reached 80% in clinically inferred MASLD-positive PermAF. The elevated FIB-4 observed in PermAF must be interpreted with explicit caution: this group was substantially older (median 79.5 years) and carried the highest burden of chronic heart failure and chronic kidney disease; therefore, in this subgroup, FIB-4 most plausibly reflects age and cardio-renal comorbidity rather than histologically confirmed hepatic fibrosis. After matching, MASLD was not an independent predictor of AF presence (OR = 0.96; 95% CI: 0.59–1.46) or its clinical severity. Conclusions: Probable MASLD, defined by clinical and non-invasive indices, was not independently associated with AF in this cohort, but AF subtypes exhibited a clear phenotypic gradient—from a metabolically driven profile in early AF to a cardio-renal and fibrotic pattern in advanced, elderly AF. Elevated FIB-4 values in PermAF most plausibly reflect age and cardio-renal comorbidity rather than true histologically confirmed hepatic fibrosis. These findings support a phenotype- and population-dependent MASLD–AF relationship and underscore the need for imaging- and histology-verified longitudinal studies. Full article

Objectives: Occupational physical activity (OPA) and leisure-time physical activity (LTPA) have contrasting health effects, a phenomenon known as the “physical activity paradox.” We explored the domain-specific associations between physical activity and non-alcoholic fatty liver disease (NAFLD). Methods: This cross-sectional study included 20,584 Korean workers (10,846 women). Physical activity was assessed using the Global Physical Activity Questionnaire, and NAFLD was assessed using the hepatic steatosis index and the presence of metabolic dysfunction. Logistic regression models were employed to explore the association between each domain of physical activity and NAFLD. The associations are presented as odds ratios (ORs) and 95% confidence intervals (CIs). Results: The prevalence of NAFLD was 30.6% in men and 18.1% in women. For male workers, ≥300 min/week of OPA was positively associated with NAFLD (OR: 1.41; 95% CI: 1.15–1.72), while ≥300 min/week of LTPA was negatively associated with NAFLD (OR: 0.79; 95% CI: 0.67–0.93). In female workers, LTPA was negatively associated with NAFLD from a lower level (OR: 0.63; 95% CI: 0.52–0.78 for 1–149 min/week; OR: 0.71; 95% CI: 0.56–0.89 for 150–299 min/week; OR: 0.58; 95% CI: 0.43–0.78 for ≥300 min/week of LTPA), while OPA had no clear association with NAFLD. Conclusions: OPA and LTPA were differentially associated with NAFLD in workers. Domain- and sex-specific effects of physical activity should be considered for the prevention and management of NAFLD. Full article

Cardiometabolic diseases are increasingly understood as disorders involving compartment-specific redox disruption rather than a uniform excess of reactive oxygen species. This narrative review synthesizes evidence for a proposed gut microbiota–mitochondria ferroptosis framework in which dysbiosis-derived lipopolysaccharide, trimethylamine N-oxide, short-chain fatty acids, bile acids, and tryptophan metabolites may modulate mitochondrial reactive species production, antioxidant defenses, iron handling, lipid peroxide detoxification, and inflammatory signaling. The reference set was assembled through searches of PubMed and Web of Science Core Collection, supplemented by targeted Google Scholar searches and citation chaining during manuscript preparation and revision through June 2026 and was organized around microbial metabolites, mitochondrial redox biology, ferroptosis pathways, disease-specific evidence, and redox-targeted interventions. Because this is a narrative synthesis rather than a systematic review, the framework should be interpreted as hypothesis-generating rather than as a systematically validated pathological model. Across atherosclerosis, diabetic cardiomyopathy, metabolic dysfunction-associated steatotic liver disease, obesity-associated insulin resistance, chronic kidney disease, and cardiorenal metabolic injury, the most consistent mechanistic links involve mtROS, impaired mitophagy, glutathione/GPX4 and SLC7A11 dysfunction, ACSL4-dependent lipid peroxidation, Nrf2 signaling, NLRP3 activation, and cGAS-STING-associated inflammation, although human causal evidence remains uneven. Importantly, much of the current literature supports local links within this sequence rather than a fully verified dysbiosis–metabolite–mitochondria ferroptosis–organ dysfunction chain in the same study. We therefore emphasize evidence tiers, terminology discipline, and biomarker requirements when interpreting ferroptosis-sensitive injury. Polyphenols, flavonoids, probiotics, postbiotics, melatonin, CoQ10-related strategies, mitochondria-targeted antioxidants, and ferroptosis-sensitive approaches may be most translatable when paired with microbiome, metabolomic, lipidomic, pharmacokinetic, and redox biomarkers. Full article

Background: MASLD is characterised by chronic inflammation and oxidative stress, which contribute to disease progression. Currently, no effective pharmacological treatment is available, and the first-line treatment remains lifestyle modification, including dietary changes and physical activity. This study aimed to assess the effect of dietary antioxidants, anthocyanins, on the risk of MASLD in a cohort from Southern Italy. Methods: The sample of this study comprised 1, 297 individuals aged between 54 and 64 years from a larger cohort, the NUTRIHEP study cohort. Data on anthocyanin intake were collected using a food-frequency questionnaire. MASLD is diagnosed when fatty liver disease is present in conjunction with at least one cardiometabolic risk factor. Results: Anthocyanin intake was inversely associated with MASLD risk. In Model b, adjusted for adjusted for age, sex, Fasting Glucose, Triglycerides, Diastolic Blood Pressure, Job, Alcohol consumption (g/day), daily energy intake, adherence to the Relative Mediterranean Diet (rMED), Available Carbohydrates, fibre intake, the third quartile (Q3) and the highest intake group (Q4) of anthocyanins showed a negative correlation with MASLD. Analysis of Anthocyanin intake as a continuous variable showed a modest negative association with MASLD risk (OR = 0.990, 95% CI 0.989–0.999), suggesting that higher anthocyanin intake may slightly lower the risk of MASLD. Conclusions: Our study highlights the protective effects of dietary anthocyanins against MASLD. These findings confirm the potential preventive role of dietary polyphenols in MASLD and identify anthocyanins as novel targets for intervention. Full article

Background/Objectives: Exposure to high and sustained levels of non-esterified fatty acids (NEFA) in the peripartal period is the main cause of fatty liver disease in dairy cows. Rumen-protected choline is often fed as part of the nutritional management of peripartal cows, with in vivo and in vitro data indicating positive effects of this nutrient on alleviating liver lipid accumulation. Although hepatic molecular mechanisms associated with choline supply have been studied using a target gene, protein, or metabolite approach, application of high-throughput technologies could vastly enhance fundamental knowledge on the functional role of choline. The main objective was to challenge isolated hepatocytes with a mixture of NEFA and determine proteome- and metabolome-wide effects in response to choline supply. Methods: Three healthy female calves (1 d old, 30–45 kg) were sacrificed to harvest hepatocytes. During a 12 h incubation, isolated hepatocytes were challenged without NEFA (control), 1.2 mM NEFA (c9-18:1, 18:2, 16:0, 18:0, and c9-16:1 at 43.5%, 4.9%, 31.9%, 14.4%, and 5.3% of total NEFA, respectively), or NEFA for 6 h followed by 10 μM choline chloride for another 6 h (NEFA + Chol). iTRAQ labeling-based protein profiling and GC/MS-based metabolomics profiling were used to determine changes in proteins and metabolites. Differentially abundant proteins for each group comparison were determined at a threshold of 1.4-fold change. Differences in metabolite profiles were assessed via pairwise comparisons. A subset of differentially abundant proteins was validated via qRT-PCR and Western blotting. Results: Compared with the control, there were 90 proteins and 22 metabolites in the NEFA group, and 83 proteins and 29 metabolites in the NEFA + Chol. Compared with NEFA, there were 49 proteins and 17 metabolites in the NEFA + Chol group. Greater abundance of hexokinase-1 (HK1), fructose-bisphosphate aldolase (ALDOA), mitochondrial pyruvate carrier 1 (MPC1), and increased concentrations of lactate with high NEFA treatment alone suggested greater glycolytic and TCA cycle activity. Accumulation of triacylglycerol in the NEFA group was associated with lipotoxicity and markers of inflammation, such as greater abundance of prostaglandin reductase 1 (PTGR1), serious cell autophagy processes, such as greater abundance of cell division cycle 42 (CDC42), and NFκB-related proteins. Choline supplementation reduced TAG partly due to greater VLDL secretion driven by greater abundance of diacylglycerol acyltransferase (DGAT1), perilipin 3 (PLIN3), and apolipoprotein C-III (APOC3). In addition, a greater abundance of carnitine O-palmitoyltransferase 1b (CPT1B) with choline suggested enhanced mitochondrial β-oxidation. Activation of the CDC42/JNK pathway and ROS/NFκB axis-related proteins, along with depressed PI3K/AKT/RAC-related proteins, indicated enhanced mitochondrial autophagy in response to NEFA. Conclusions: Overall, data confirmed published effects of choline on TAG accumulation, VLDL secretion, and fatty acid oxidation, while highlighting negative effects of NEFA on the respiratory electron transport chain, autophagy, and inflammatory processes. Full article