Metabolic dysfunction-Associated Steatotic Liver Disease (MASLD) and associated dyslipidemia is a growing health issue that gives rise to cardiovascular risk. Men are more prone to development of MASLD than women. Understanding mechanisms underlying sex differences in MASLD may lead to improved prevention and treatment approaches. Cholesteryl ester transfer protein (CETP) is a lipid transfer protein that shuttles triglycerides and cholesteryl esters between blood lipoproteins and tissues. In this study investigate the impact of hepatic CETP expression on MASLD. Hepatic CETP expression (L-HuCETP) was achieved by injecting liver-targeted CETP-expressing adeno-associated virus into C57BL/6J mice. In females, L-HuCETP improved glucose tolerance, consistent with our prior clamp results in global human CETP transgenic mice. Whereas in males, L-HuCETP worsened glucose metabolism and impaired insulin signaling. Correspondingly, L-HuCETP expression reduced the expression of gluconeogenic pathway genes in females but upregulated these genes in males. In males, L-HuCETP mice exhibited increased hepatic lipid droplet accumulation, lipogenesis proteins and these changes were not observed in females. L-HuCETP expression resulted in sex-specific hepatic responses, with increased expression of inflammation and fibrosis related genes in male, but decreased expression of these genes in females. Mechanistic studies indicate that L-HuCETP had sex specific effects on transcription factors ChREBP and HNF4α, which are important for glucose and lipid metabolism. Our studies suggest that sex-specific roles of L-HuCETP with regard to liver metabolic adaptation and MASLD risk in obesity, highlighting CETP-mediated pathways as potential targets for sex-specific precision medicine approaches to improve MASLD.

Food-derived bioactive components play a crucial role in the prevention of cardiovascular diseases. Daidzein (DAI), a soy isoflavone, possesses notable cardioprotective potential; however, its precise role in nicotine-induced atherosclerosis (AS) and its underlying mechanisms remain unclear. This study explored whether DAI protects against AS by modulating macrophage pyroptosis and lipid metabolism via redox-sensitive signaling pathways. In human acute monocytic leukemia cell line (THP-1)-derived macrophages, DAI alleviated nicotine-induced pyroptosis, as evidenced by the downregulation of NOD-like receptor family pyrin domain-containing 3 (NLRP3), ASC, cleaved caspase-1, and gasdermin D (GSDMD)-N, accompanied by reduced secretion of interleukin (IL)-1β, IL-6, and IL-18, and decreased lactate dehydrogenase (LDH) release and caspase-1 activity. DAI also enhanced (7-nitrobenz-2-oxa-1,3-diazole)-cholesterol (NBD)-cholesterol efflux, suppressed DiI-labeled oxidized low-density lipoprotein (DiI-ox-LDL) uptake, and mitigated intracellular lipid droplet accumulation. Mechanistically, silencing nuclear factor erythroid 2-related factor 2 (Nrf2), inducing reactive oxygen species (ROS), or overexpressing thioredoxin-interacting protein (TXNIP) abolished the protective effects of DAI, confirming the involvement of the Nrf2/ROS/TXNIP pathway. In nicotine-exposed apolipoprotein E-deficient (apoE-/-) mice, DAI upregulated vascular Nrf2, downregulated TXNIP and pyroptosis-related proteins, improved plasma lipid profiles, alleviated systemic inflammation, and reduced aortic plaque formation, with no obvious adverse effects observed under the experimental conditions used. Collectively, DAI attenuated macrophage pyroptosis and lipid accumulation via the Nrf2/ROS/TXNIP pathway, thereby mitigating nicotine-induced atherogenesis. These findings provide preclinical evidence for the potential of DAI as a food-derived bioactive compound for smoking-related AS.

Long noncoding RNA small nucleolar RNA host gene 5 (SNHG5) has been implicated in cell death, glucose homeostasis, and tumor progression, yet its role in atherosclerosis (AS) remains unclear. In this study, SNHG5 expression was markedly elevated in aortic tissues of high-fat diet-fed apoE-/- mice and in ox-LDL-stimulated THP-1 macrophages. Lentiviral-mediated SNHG5 silencing in vivo reduced plaque burden, attenuated lipid deposition, increased collagen content, and decreased systemic inflammation. Moreover, SNHG5 knockdown increased plasma HDL-C and promoted reverse cholesterol transport. In mouse peritoneal macrophages and THP-1-derived foam cells, SNHG5 silencing enhanced cholesterol efflux to lipid-free apoA-I without affecting uptake, accompanied by upregulation of ABCA1. LXRα expression remained unchanged, whereas HDAC3 was downregulated; HDAC3 overexpression reversed the effects of SNHG5 knockdown on ABCA1 expression, cholesterol efflux, and foam cell formation. Subcellular fractionation indicated cytoplasmic localization of SNHG5, and luciferase reporter and RNA pull-down assays confirmed that it functions as a competing endogenous RNA for miR-216a-5p. Inhibition of miR-216a-5p largely abolished the effects of SNHG5 silencing, establishing the SNHG5/miR-216a-5p/HDAC3 axis as a key regulator of macrophage lipid handling. Collectively, these findings demonstrate that SNHG5 promotes macrophage lipid accumulation and atherogenesis by sequestering miR-216a-5p to upregulate HDAC3 and suppress ABCA1-mediated cholesterol efflux, highlighting SNHG5 as a potential therapeutic target for AS.

C57BL/6J mice are widely used in biomedical research and are susceptible to insulin resistance and dyslipidemia when challenged with high fat diets relative to other inbred strains. Interestingly, C57Bl/6J mice contain a naturally occurring premature termination codon in the lipid flippase Atp10D, and previous studies have linked Atp10D to the metabolic disease-prone phenotype in mice and atherosclerotic severity in humans. In this study, we used CRISPR/Cas9 to revert the premature termination codon to the wild-type glutamine codon (Atp10D *817Q) in the C57Bl/6J mouse strain. The RNA transcripts from original and corrected alleles are dually expressed in heterozygous mice, suggesting that the mutant transcript escapes nonsense-mediated decay. Expression of two corrected Atp10D alleles restores wildtype expression levels of the transcript and protein in the liver. When challenged with a high fat diet, Atp10D-/- (original) and Atp10D+/+ (corrected) C57Bl/6J mice showed no significant difference in weight gain, glucose tolerance, or plasma levels of triglycerides, cholesterol, or free fatty acids. However, the female Atp10D+/+ mice displayed an increase in complex glycosphingolipids and a reduction in cardiolipins in the plasma. These results suggest that restoring the expression of Atp10D in C57Bl/6J mice does not reverse insulin resistance and dyslipidemia in response to high fat diet feeding.

Background:Hormone therapy (HT) has not consistently reduced atherosclerotic cardiovascular disease (ASCVD) events in post-menopausal women, yet the underlying mechanisms remain poorly understood. Methods:Female Ldlr -/- mice with established atherosclerosis were subjected to surgical menopause and treated with 17β-estradiol (E2) following lipid normalization. Studies were performed in aging and young mice. To determine whether inflammation mediates the age-dependent response to HT, a cohort of aging mice underwent transplantation with Ifnγ -/- bone marrow (BM) before hormone treatments. Metabolic parameters, HDL function, systemic inflammation, atherosclerotic burden, liver metabolic and oxidative stress signaling, and hepatic estrogen receptor signaling were evaluated. Results:In aging mice, menopause E2 treatment failed to reduce established atherosclerosis as shown in sham operated mice during lipid normalization. Instead, E2 treatment increased circulating IFNγ and IL-6, impaired HDL antioxidant and cholesterol efflux functions, and promoted inflammatory and vulnerable plaque phenotypes. Suppression of inflammation through Ifnγ -/-BM transplantation restored HDL function and significantly reduced atherosclerosis in E2-treated aging mice. In contrast to aging mice, young mice exhibited reduced systemic and plaque inflammation, improved HDL functions and atherosclerosis following E2 treatment. Liver RNA sequencing and qPCR validation identified activation of inflammatory, oxidative stress, and lipid metabolic pathways in aging E2-treated mice, which were largely attenuated following Ifnγ -/- bone marrow transplantation as well as in young mice. Compared to young mice, aging mice presented hepatic estrogen receptor remodeling characterized by reduced estrogen receptor α (ERα) expression and increased G-protein coupled estrogen receptor (GPER) expression. Constitutive GPER activation was accompanied by induction of NOX1-dependent oxidative stress, which was further exacerbated by E2 treatment, leading to persistent inflammation. Conclusions:The cardiovascular effects of estrogen therapy are fundamentally age dependent. Aging shifts estrogen signaling toward hepatic oxidative stress and inflammation through increased GPER. While E2 treatment preserves both metabolic and cardiovascular protection in young mice, aging exacerbates GPER-NOX1-mediated oxidative stress, resulting in impaired HDL function and persistent residual ASCVD risk. These findings identify inflammation-driven, non-lipid mechanisms as potential therapeutic targets to improve cardiovascular outcomes during hormone therapy in postmenopausal women.

Primary hepatocellular carcinoma (PHC) is the sixth most common cancer and the third leading cause of cancer death worldwide. Hepatocellular carcinoma (HCC) accounts for 75%-85% of PHC. LARP3 is aberrantly expressed in multiple cancers. We found that it is significantly highly expressed in the liver cancer tissues of HCC patients, but the exact role and specific mechanism of this abnormal expression are not yet clear. In this study, through bioinformatics analysis, we concluded that LARP3 expression is associated with a poor prognosis for patients with HCC. Through cellular experiments such as gene editing and phenotypic functions, we found that LARP3 promotes the occurrence and development of HCC and inhibits apoptosis. Finally, through biological means such as RNA sequencing, flow cytometry, western blotting, and the construction of a subcutaneous tumorigenesis model in nude mice, we concluded that inhibition of HCC apoptosis by LARP3 is related to LARP3 negatively regulating ROS level and inhibiting the PI3K/c-Fos/apoptosis axis. This study will provide potential targets for the treatment of HCC.

Background/Objectives: Studies have shown that Atractylenolide I (AT-I) can exert anti-inflammatory and anti-oxidative effects, protecting against the development of various kinds of cardiovascular diseases. However, whether AT-I prevents nicotine-induced atherogenesis is unknown. This study was designed to explore the effects of AT-I on nicotine-induced macrophage pyroptosis and the progression of atherosclerosis. Methods: RT-qPCR and Western blot were used to detect the mRNA and protein levels of TXNIP and pyroptosis-related factors in THP-1-derived macrophages. ELISA was used to detect the secretion of pro-inflammatory cytokines. Hoechst/PI double-staining assay was used to assess plasma membrane integrity. The ROS assay kit, LDH release assay kit, and caspase-1 activity assay kit were used to detect ROS production, LDH release, and caspase-1 activity. Oil Red O, HE, and Masson staining were used to evaluate lipid accumulation, lesion size, and plaque stability in HFD-fed apoE-/- mice. Results: AT-I treatment significantly decreased pyroptosis-related factors expression, disrupted plasma membrane integrity, and down-regulated pro-inflammatory cytokines secretion, thereby inhibiting nicotine-induced pyroptosis of THP-1-derived macrophages. In addition, AT-I decreased ROS production and the expression of TLR4 and TXNIP. Lentivirus overexpression of TLR4 or TXNIP, or pre-treatment with ROS agonist, mainly reversed the anti-pyroptotic effects of AT-I in nicotine-treated THP-1-derived macrophages. Additionally, administering AT-I to HFD-fed apoE-/- mice markedly decreased nicotine-induced up-regulation of pyroptosis-related proteins in the aortas. Enzymatic methods and ELISA assay suggested that AT-I improved dyslipidemia and inflammation in vivo. Oil Red O, HE, and Masson staining showed that AT-I alleviated lipid accumulation, decreased plaque size, and increased plaque stability. Conclusions: Taken together, AT-I can be regarded as a potential phytomedicine that protects against macrophage pyroptosis and atherosclerosis triggered by nicotine.

Atherosclerosis is a chronic inflammatory disease characterized by lipid accumulation, immune dysregulation, and cell death within the arterial wall. While extensive research has been devoted to the understanding of its molecular pathogenesis, novel regulatory factors contributing to its progression remain to be identified. Macrophage pyroptosis and the subsequent inflammatory response play a central role in atherogenesis. Recent evidence has implicated long non-coding RNAs (lncRNAs) in macrophage dysfunction and vascular inflammation; however, the precise roles of specific lncRNAs in pyroptosis-mediated atherogenesis are still unclear. In this study, we identified lncRNA MIR17HG as a potential regulator of macrophage pyroptosis and atherosclerosis progression. Exposure of THP-1-derived macrophages to ox-LDL induced a dose-dependent increase in MIR17HG expression. Silencing of MIR17HG significantly reduced the expression of pyroptosis-related factors, caspase-1 activity, LDH release, proinflammatory cytokine secretion, and plasma membrane disruption in ox-LDL-treated macrophages, indicating attenuation of pyroptosis. These effects were reversed by TXNIP overexpression or transfection with a miR-301a-3p inhibitor. Mechanistically, MIR17HG functioned as a competing endogenous RNA (ceRNA), sponging miR-301a-3p to upregulate TXNIP expression and activate the NLRP3 inflammasome. Consistently, MIR17HG knockdown reduced plaque burden, improved plasma lipid profile, and alleviated inflammation in high-fat diet-fed apoE-/- mice; these protective effects were abrogated by administration of a miR-301a-3p antagomir. Collectively, our findings reveal that MIR17HG promotes atherosclerosis by enhancing macrophage pyroptosis through the miR-301a-3p/TXNIP/NLRP3 axis, offering new insight into the lncRNA-mediated regulation of vascular inflammation and plaque development.

Understanding how exercise training or a healthy diet improves metabolic outcomes is crucial to developing therapeutic targets for obesity-associated diseases. In this study, obesity was induced by a 12-week high-fat diet (HFD) feeding in male and female mice. Then, mice of each sex were divided into 3 groups of lifestyle interventions that followed for 6 weeks: a. HFD; b. chow diet; c. HFD+exercise; d. chow diet +exercise. Age-matched mice continuously fed a chow diet served as controls. An oral glucose tolerance test was performed 48 hours after the final exercise session. HFD increased body weight, fasting blood triacylglycerol (TG) and insulin levels only in male mice, while increased body fat composition, fasting glucose and cholesterol, hepatic steatosis, and impaired glucose tolerance in both sexes. Glucose tolerance was improved in both sexes by all three interventions: the chow diet, exercise training, or the combination of both. However, sex differences were presented in improving certain pathways in the liver or skeletal muscle tissues in response to the chow diet or exercise training. Furthermore, the chow diet reduced body fat composition, hepatic steatosis, and fasting cholesterol in both sexes. Exercise training reduced body fat and fasting lipid levels only in male mice and appeared more beneficial to males than females in improving glucose metabolism. Regarding HDL functions, exercise training improved the antioxidant activity of HDL particles in both lean and obese mice of both sexes, independent of weight-loss or diet. There was no combined effect of diet and exercise training in improving glucose tolerance once body weight is corrected to control levels by either diet or exercise. Further analysis of molecular pathways demonstrated several gene sets modified by HFD were reversed by all three lifestyle interventions, which were tissue specific, either in the liver or skeletal muscles. Results from your studies may provide insight into the metabolic pathways that could be potentially targeted for improving metabolic diseases. L Zhu (K01AG077038); J Stafford (BX002223) (R01DK109102) (R01HL144846). This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.

IntroductionHigh density lipoproteins (HDL) exert cardiovascular protection in part through their antioxidant capacity and cholesterol efflux function. Effects of exercise training on HDL function are yet to be well established, while impact on triacylglycerol (TG)-lowering has been often reported. We previously showed that a short-term high-intensity interval training (HIIT) program improves insulin sensitivity but does not inhibit inflammatory pathways in immune cells in insulin-resistant subjects. The purpose of this study is to evaluate HDL function along with changes of lipoproteins after the short-term HIIT program in lean, obese nondiabetic, and obese type 2 diabetic (T2DM) subjects.MethodsAll individuals underwent a supervised 15-day program of alternative HIIT for 40 minutes per day. VO2peak was determined before and after this training program. A pre-training fasting blood sample was collected, and the post-training fasting blood sample collection was performed 36 hours after the last exercise session.ResultsBlood lipid profile and HDL function were analyzed before and after the HIIT program. Along with improved blood lipid profiles in obese and T2DM subjects, the HIIT program affected circulating apolipoprotein amounts differently. The HIIT program increased HDL-cholesterol levels and improved the cholesterol efflux capacity only in lean subjects. Furthermore, the HIIT program improved the antioxidant capacity of HDL in all subjects. Data from multiple logistic regression analysis showed that changes in HDL antioxidant capacity were inversely associated with changes in atherogenic lipids and changes in HDL-TG content.DiscussionWe show that a short-term HIIT program improves aspects of HDL function depending on metabolic contexts, which correlates with improvements in blood lipid profile. Our results demonstrate that TG content in HDL particles may play a negative role in the anti-atherogenic function of HDL.

Genetic association studies have linked ATP10A and closely related type IV P-type ATPases (P4-ATPases) to insulin resistance and vascular complications, such as atherosclerosis. ATP10A translocates phosphatidylcholine and glucosylceramide across cell membranes, and these lipids or their metabolites play important roles in signal transduction pathways regulating metabolism. However, the influence of ATP10A on lipid metabolism in mice has not been explored. Here, we generated gene-specific Atp10A knockout mice and show that Atp10A-/- mice fed a high-fat diet did not gain excess weight relative to wild-type littermates. However, Atp10A-/- mice displayed female-specific dyslipidemia characterized by elevated plasma triglycerides, free fatty acids and cholesterol, as well as altered VLDL and HDL properties. We also observed increased circulating levels of several sphingolipid species along with reduced levels of eicosanoids and bile acids. The Atp10A-/- mice also displayed hepatic insulin resistance without perturbations to whole-body glucose homeostasis. Thus, ATP10A has a sex-specific role in regulating plasma lipid composition and maintaining hepatic liver insulin sensitivity in mice.

Ferroptosis is a newly identified form of non-apoptotic programmed cell death, characterized by the iron-dependent accumulation of lethal lipid reactive oxygen species (ROS) and peroxidation of membrane polyunsaturated fatty acid phospholipids (PUFA-PLs). Ferroptosis is unique among other cell death modalities in many aspects. It is initiated by excessive oxidative damage due to iron overload and lipid peroxidation and compromised antioxidant defense systems, including the system Xc-/ glutathione (GSH)/glutathione peroxidase 4 (GPX4) pathway and the GPX4-independent pathways. In the past ten years, ferroptosis was reported to play a critical role in the pathogenesis of various cardiovascular diseases, e.g., atherosclerosis (AS), arrhythmia, heart failure, diabetic cardiomyopathy, and myocardial ischemia-reperfusion injury. Studies have identified dysfunctional iron metabolism and abnormal expression profiles of ferroptosis-related factors, including iron, GSH, GPX4, ferroportin (FPN), and SLC7A11 (xCT), as critical indicators for atherogenesis. Moreover, ferroptosis in plaque cells, i.e., vascular endothelial cell (VEC), macrophage, and vascular smooth muscle cell (VSMC), positively correlate with atherosclerotic plaque development. Many macromolecules, drugs, Chinese herbs, and food extracts can inhibit the atherogenic process by suppressing the ferroptosis of plaque cells. In contrast, some ferroptosis inducers have significant pro-atherogenic effects. However, the mechanisms through which ferroptosis affects the progression of AS still need to be well-known. This review summarizes the molecular mechanisms of ferroptosis and their emerging role in AS, aimed at providing novel, promising druggable targets for anti-AS therapy.

Background: Metabolic dysfunction -associated steatotic liver disease (MASLD) is a common complication of obesity and, in severe cases, progresses to metabolic dysfunction -associated steatohepatitis (MASH). Small heterodimer partner (SHP) is an orphan member of the nuclear receptor superfamily and regulates metabolism and inflammation in the liver via a variety of pathways. In this study, we investigate the molecular foundation of MASH progression in mice with hepatic SHP deletion and explore possible therapeutic means to reduce MASH. Methods: Hepatic SHP knockout mice (SHPDhep) and their wild -type littermates (SHPfl/fl) of both sexes were fed a fructose diet for 14 weeks and subjected to an oral glucose tolerance test. Then, plasma lipids were determined, and liver lipid metabolism and inflammation pathways were analyzed with immunoblotting, RNAseq, and qPCR assays. To explore possible therapeutic intersections of SHP and inflammatory pathways, SHPDhep mice were reconstituted with bone marrow lacking interferon y (IFNy-/-) to suppress inflammation. Results: Hepatic deletion of SHP in mice fed a fructose diet decreased liver fat and increased proteins for fatty acid oxidation and liver lipid uptake, including UCP1, CPT1a, ACDAM, and SRBI. Despite lower liver fat, hepatic SHP deletion increased liver inflammatory F4/80+ cells and mRNA levels of inflammatory cytokines (IL -12, IL -6, Ccl2, and IFNy) in both sexes and elevated endoplasmic reticulum stress markers of Cox2 and CHOP in female mice. Liver bulk RNAseq data showed upregulation of genes whose protein products regulate lipid transport, fatty acid oxidation, and inflammation in SHPDhep mice. The increased inflammation and fibrosis in SHPDhep mice were corrected with bone marrow -derived IFNy-/- myeloid cell transplantation. Conclusion: Hepatic deletion of SHP improves fatty liver but worsens hepatic inflammation possibly by driving excess fatty acid oxidation, which is corrected by deletion of IFNy specifically in myeloid cells. This suggests that hepatic SHP limits fatty acid oxidation during fructose diet feeding but, in doing so, prevents pro -MASH pathways. The IFNy-mediated inflammation in myeloid cells appears to be a potential therapeutic target to suppress MASH. Published by Elsevier GmbH. This is an open access article under the CC BY -NC -ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Premenopausal women and endurance-trained individuals of either sex have reduced cardiovascular disease (CVD) risk. Endurance training shifts fuel selection towards fats to spare carbohydrates; interestingly, women prioritize fats as an energy resource more than men do during exercise. Relying on fats during exercise drives whole-body lipolysis and promotes lipid uptake and oxidation capacity in skeletal muscles. These metabolic adaptations during exercise result in protection against diet-induced obesity, a healthy body fat distribution, and reduced plasma triacylglycerol (TG) concentrations. Here, we analyze how sex differences and endurance training mediate changes in skeletal muscles, including exercise-induced lipolysis, lipid uptake and β-oxidation, intramuscular TG storage, and postexercise lipid metabolism, and discuss how regulating this processes affects CVD risk.

Some polysaccharides with established medical and nutritional values have been identified to possess immunomodulatory properties devoid of any toxic or adverse effects. Previous studies have demonstrated that water-soaked polysaccharides from the skin of white asparagus can enhance cytokine release in RAW 264.7 macrophages, however, the underlying mechanism governing immune regulation remains elusive. In this study, we obtained a lower molecular weight polysaccharide (AP) through acid extraction, with an average MW of approximately 9.5 kDa. SEM and AFM spectroscopy analysis revealed well-dispersed spherical particle with triple helix conformation for AP, characterized by intertwined branching structures. Treatment with AP resulted in a time-dependent increase in nitric oxide levels and cytokine production in both RAW 264.7 cells and primary peritoneal macrophages. RNA-seq analysis indicated that AP activated macrophages via NLRP3 inflammasome signaling pathway. Furthermore, AP activated MAPKs and JAK/STAT signaling pathways to amplify the inflammatory response. Additionally, administration of AP improved visceral index and reduced inflammatory cell counts in CYP-induced immunosuppressed mice models. These findings suggest that AP holds potential as an immuno-enhancement mediator, wherein MAPK and JAK/STAT3 signaling pathways play a role in NLRP3 inflammasome activation of macrophages.