High-fat High-cholesterol Diet Research Articles (Page 1)

Background: Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) has become a worldwide health concern. Soy protein isolate (SPI) is a plant-derived protein with high nutritional value and has shown promising effects in regulating lipid metabolism and inflammation. Objectives: This study aimed to investigate the effects of an α-subunit-deficient SPI (α-lack-SPI) on MASLD and the underlying molecular mechanisms. Methods: Rats were fed with a high-fat, high-cholesterol diet (HFD) to induce MASLD. Results: The results showed that α-lack-SPI significantly reduced the levels of hepatic TG and TC, serum ALT, AST, TC, and LDL-C, and increased serum HDL-C in rats with HFD-induced MASLD. α-lack-SPI significantly attenuated hepatic steatosis and hepatocyte ballooning revealed by histopathological analysis. Meanwhile, α-lack-SPI markedly downregulated the mRNA expressions of Srebf1, Acaca, Fasn, Pcsk9, and Hmgcr, while significantly upregulating Pparα. Additionally, α-lack-SPI treatment significantly reduced the mRNA expressions of hepatic pro-inflammatory cytokines (Tnf-α, Il-1β, Il6), chemokine (Ccl2), and inflammasome component (Nlrp3), as well as the protein expression of COX-2. Conclusions: In conclusion, α-lack-SPI alleviated MASLD in HFD-fed rats probably via improving hepatic lipid metabolism and mitigating hepatic inflammation. These findings indicate that α-lack-SPI may serve as a promising nutritional intervention for MASLD management.

The prevalence of hyperuricemia with elevated serum uric acid is increasing worldwide. However, the effects of high uric acid on diabetic patients with dyslipidemia and the mechanisms underlying these effects remain unexplored. This study aimed to develop a novel diabetic model of hyperuricemia and dyslipidemia in male hamsters to evaluate the effects of high uric acid on glucolipid metabolism, renal injury and the gut microbiota. Twelve healthy hamsters were randomly divided into two groups and fed with a normal diet and high-fat/cholesterol diet (HFCD), respectively. Twenty-four diabetic hamsters were randomly divided into four groups receiving a normal diet; HFCD; potassium oxonate (PO) treatment (intragastric PO at doses of 350 mg/kg and adenine at doses of 150 mg/kg with 5% fructose water); and PO treatment with HFCD, respectively. After 4 weeks, all animals were dissected for determining serum biochemical indicators, tissue antioxidant parameters, renal pathological changes, target gene expressions, fecal short-chain fatty acids content, and the gut microbiota composition. The results showed that a hamster model with hyperuricemia and dyslipidemia was successively established by the combination of PO treatment and HFCD, in which serum uric acid, glucose, triglyceride and total cholesterol levels reached 499.5 ± 61.96 μmol/L, 16.88 ± 2.81 mmol/L, 119.88 ± 27.14 mmol/L and 72.92 ± 16.62 mmol/L, respectively. PO treatment and HFCD had synergistic effects on increasing uric acid, urea nitrogen, creatinine levels, liver xanthine oxidase activity, plasminogen activator inhibitor-1 and transforming growth factor-β expressions, and the relative abundance of Lleibacterium (p < 0.05); in addition, they caused glomerular mesangial cells and matrix proliferation, protein casts and urate deposition. High uric acid was closely related to decreased antioxidant capacity; decreased renal vascular endothelial growth factor expression; increased acetic acid content; decreased butyric, propanoic, and isobutyric acid levels; decreased Firmicutes to Bacteroidetes ratios (p < 0.05); and altered epithelial integrity and structure of the gut microbiota in diabetic hamsters. The findings indicate that high uric acid affects the glucolipid metabolism, accelerates renal damage, and disrupts the balance of intestinal flora in diabetic animals, which provides a scientific basis for metabolic syndrome prevention and control in diabetes.

Oxidative stress (OS), inflammation, and pyroptosis are hallmarks of atherosclerosis pathogenesis. Thioredoxin-interacting protein (TXNIP) crosslinks them through activating nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3) inflammasome pathway. The current work addresses the potentials of empagliflozin (EMPA) on OS markers, thioredoxin (TRX), and TXNIP, downstream mediators of NLRP3 inflammasome and the executioner of pyroptosis; gasdermin D (GSDMD) in a model of atherosclerosis. Atherosclerosis was induced by high fat cholesterol diet (HFCD) and streptozotocin (STZ) in male Wistar rats. Five groups were allocated: Control group, HFCD-STZ group, and three orally EMPA treated groups with HFCD-STZ in doses (2.5, 5, and 10mg/kg/d for 4weeks). To access the underlying mechanism of EMPA on atherosclerosis, serum glycated haemoglobin, lipid profile, inflammatory cytokines, and aortic OS markers were evaluated. Polymerase chain reaction analysis of NLRP3 inflammasome components, TRX, TXNIP, GSDMD, and vascular cell adhesion molecule-1 (VCAM-1) mRNA expressions were performed. Histopathological studies further confirmed EMPA's effects on atherosclerosis. EMPA ameliorates metabolic and histopathological alterations, inflammation, OS, TRRX/TXNIP reversal, VCAM-1, activation of NLRP3 inflammasome, and pyroptosis axis. EMPA displayed antioxidant and anti-inflammatory effects through attenuating the OS/TXNIP/NLRP3/GSDMD axis in an experimental model of atherosclerosis.

High fat and high cholesterol diet induced cognitive impairment in GSDMD knockout mice via the compensatory activation of apoptosis pathway.

A review of recent publications demonstrating the important role of the intestine in the pathogenesis of cardiovascular disease. At baseline (≥ 6 months since myocardial infarction), the pattern of fecal microbiota and blood LPS levels after a meal predicted risk for new major adverse cardiovascular events over the next 7 years. In intestine, tryptophan is metabolized primarily by the kynurenine pathway, which is regulated by the enzyme indoleamine-2,3-dioxygenase 1 (IDO1). Tryptophan flow into the kynurenine pathway limits its availability for the formation of microbiota-derived indole derivatives including butyrate, and limits availability of tryptophan for the 5-hydroxytryptamine (5-HT or serotonin) pathway. Feeding Ldlr-/- mice a high-fat high-cholesterol diet (HFD+HCD) increased intestinal IDO1 activity, decreased levels of tryptophan, fecal butyrate, and 5-HT. Ldlr-/- mice deficient in intestinal Ido1 ( Ldlr-/-Ido1-/- ) on HFD+HCD had increased intestinal levels of 5-HT, increased gut permeability, increased gut inflammation, increased LPS, and increased aortic atherosclerosis. Ldlr-/- mice fed HFD+HCD and treated with a 5-HT pathway inhibitor had increased fecal indole levels, improved gut-barrier, increased antimicrobial peptide levels, and decreased aortic atherosclerosis without a change in plasma cholesterol. These studies demonstrate the importance of microbiota-derived products and intestinal tryptophan metabolism in atherosclerosis.

Background/Objectives: Apolipoprotein E receptor-2 (apoER2) exists in various alternatively spliced forms, including variants that express apoER2 with or without exon 19 in the cytoplasmic domain. This study compared vascular response to endothelial denudation, as well as diet-induced atherosclerotic and metabolic diseases, between genetically modified mice that exclusively expressed the apoER2 splice variant with or without exon 19 to determine the impact of apoER2 exon 19 motif in cardiometabolic disease modulation. Methods: Vascular response to injury was assessed by measuring neointima area of the carotid arteries after endothelial denudation. The genetically modified mice were also fed a high-fat high-cholesterol diet for 16 weeks for the determination of body weight gain, glucose and insulin levels, glucose tolerance and insulin secretion. Additionally, adipose tissue inflammation was assessed by analysis of adipose gene expression, and atherosclerosis was characterized by measuring fatty lesion size in the whole aorta, as well as in the aortic roots. Results: The results showed that whereas the expression of either splice variant is sufficient to impede denudation-induced fibrotic neointima formation and complex necrotic atherosclerotic lesions, the expression of the apoER2 splice variant containing exon 19 is necessary for the complete protection of injury-induced neointima formation in the vessel wall. However, exclusive expression of either apoER2 cytoplasmic splice variant does not influence the early phase of atherogenesis. Additionally, the exclusive expression of apoER2 without exon 19 promotes adipocyte inflammation and accelerates diet-induced insulin resistance and glucose intolerance. Conclusions: These results indicate that the apoER2 cytoplasmic variants have distinct and cell type-specific roles in influencing cardiometabolic disease development.

Hepatoprotective drug screening identifies daclatasvir, a promising therapeutic candidate for MASLD by targeting PLIN2.

Nonalcoholic fatty liver disease, a progressive condition caused by the accumulation of fat in the liver, begins with simple steatosis and can potentially progress to metabolic dysfunction-associated steatohepatitis (MASH) in the presence of inflammation and fibrosis, ultimately leading to cirrhosis or hepatocellular carcinoma. Increasing evidence indicates that sodiumglucose cotransporter 2 (SGLT2) inhibitors effectively alleviate MASH in mouse models. However, there is a lack of research on the effects of enavogliflozin on liver disease. In the present study, we investigated the effects of SGLT2 inhibitors on MASH induced by a high-fat, high-cholesterol (HFHC) diet in mice. Male C57BL/6 mice were fed a normal chow diet, HFHC diet, or HFHC diet with enavogliflozin for 12 weeks. LX-2 and HepG2 cells were treated with enavogliflozin in the presence of various pathological stimuli. The HFHC diet induced excessive hepatic lipid accumulation, inflammation, and severe fibrosis. Administration of enavogliflozin not only ameliorated hepatic steatosis and fibrotic conditions but also suppressed the production of inflammatory cytokines. Positive outcomes were also observed in in vitro experiments, where enavogliflozin demonstrated the ability to impede the activation of hepatic stellate cells and alleviate lipid accumulation in hepatocytes. The potential pathway through which enavogliflozin attenuated liver fibrosis development may be associated with the transforming growth factor β1/Smad signaling pathway. Our results suggest that enavogliflozin is effective in a mouse model of MASH by attenuating hepatic steatosis, suppressing inflammation, and improving liver fibrosis.

Atherosclerosis (AS), a common cardiovascular condition, is often linked to cognitive dysfunction. This study investigates how aerobic exercise affects cognitive impairments caused by AS in ApoE-/- mice. At 8 weeks old, male ApoE-/- mice were fed a high-fat, high-cholesterol diet (HFHCD) for 6 weeks to induce AS, with C57BL/6J mice on a standard diet as control group (WT). Initially, the study compared aortic plaque and cognitive performance between the WT and AS mice. Then, AS mice were divided into sedentary (AS-SED) and exercise (AS-EX) groups for a 10-week aerobic exercise intervention. Results showed significant aortic plaques and cognitive deficits in AS mice after 6 weeks on the HFHCD diet. However, the 10-week exercise reduced plaque, improved cognition, and enhanced cerebral blood flow. Exercise intervention also decreased IL-33 expression in the hippocampus and inhibited NF-κB and IκBα phosphorylation. Furthermore, aerobic exercise reduces M1 microglial activation and pro-inflammatory cytokines like IL-6, TNF-α, and IL-1β in the hippocampus of AS mice, thereby decreasing neuroinflammation. In summary, aerobic exercise can effectively improve cognitive function by decreasing IL-33 expression and inhibiting NF-κB activation, which in turn reduces microglial activation and the release of inflammatory mediators in the hippocampus. This study provides evidence for aerobic exercise to improve cognitive impairment caused by AS.

Alpha-aminobutyric acid (ABA) is a nonproteinogenic amino acid, a metabolite which could be generated from the metabolism of methionine, threonine, serine and glycine or as a gut-microbiome-derived metabolite. Changes in ABA levels have been embroiled in metabolic dysfunction-associated steatotic liver disease (MASLD) intervention studies, but their relation to MASLD pathogenesis remains unclear. Hence, this present study aimed to investigate the effect of oral ABA supplementation on the progression of a high fat/high cholesterol diet (HFD) induced MASLD mice model. ABA was found to remodel the gut microbiome composition and ameliorate MASLD parameters in HFD-fed mice. ABA mitigated HFD-induced gain in liver weight, hepatic steatosis, insulin resistance, serum and hepatic triglyceride levels, and liver cholesterol levels. Modulation of lipid metabolism was observed in the liver, in which expression of proteins and/or genes involved in de novo lipogenesis were suppressed, while those involved in fatty acid oxidation and autophagy were upregulated together with cellular antioxidant capacity, in addition to the enhancement of the AMPK/SIRT1 pathway. ABA reshaped the gut composition by enriching nine bacterial species, including Helicobacter hepaticus, Desulfovibrio sp. G11, Parabacteroides distasonis, and Bacteroides fragilis, while diminishing the abundance of 16 species, which included four Helicobacter species. KEGG pathway analysis of microbial functions found that ABA impeded secondary bile acid biosynthesis - which was reflected in the faecal BA composition analysis. Notably, ABA also inhibited ileal FXR-Fgf15 signaling, allowing for increased hepatic Cyp7a1 expression to eliminate cholesterol buildup in the liver. Overall, our findings indicate that ABA could be used as a promising therapeutic approach for the intervention of MASLD.

Invivo base editing of Angptl3 via lipid nanoparticles to treat cardiovascular disease.

Silicon-enriched meat consumption mitigates brain cortex damage associated with diabetic dyslipidemia in a late-stage type 2 diabetes mellitus rat model

Background and AimsMetabolic-associated steatohepatitis (MASH) is an advanced and progressive liver disease that potentially causes cirrhosis and hepatocellular carcinoma. Exercise is a crucial and effective intervention for ameliorating metabolic dysfunction-associated steatotic liver disease. This study aimed to provide a comprehensive understanding of the underlying mechanisms of MASH, which benefit a broad spectrum of MASH patients, including those who have difficulty engaging in physical activity.MethodsWe established a mouse model of MASH and selectively knocked down L-type amino acid transporter 1 and alanine-serine-cysteine transporter 2. Mice were fed a high-fat high-cholesterol diet and subjected to either short- or long-term exercise regimens. We assessed the phosphorylation and activity of branched-chain alpha-keto acid dehydrogenase (BCKDH) as well as branched-chain amino acid (BCAA) content in skeletal muscle following exercise.ResultsShort-term exercise significantly reduced hepatic steatosis and inflammation without causing notable changes in body weight. It also enhanced BCKDH activity in skeletal muscle and decreased hepatic BCAA accumulation. Muscle-specific overexpression of BCKDH further promoted BCAA catabolism and significantly attenuated hepatic steatosis and inflammation in high-fat high-cholesterol-fed mice. In contrast, muscle-specific L-type amino acid transporter 1 knockdown, which suppresses BCAA uptake, markedly abolished these beneficial effects. Interestingly, BCKDH overexpression in muscle increased glutamine levels in both the blood and liver. Hepatic alanine-serine-cysteine transporter 2 knockdown, which inhibited glutamine uptake, lessened the protective effect of exercise on MASH. Further in vitro study revealed that glutamine derived from myocytes improved redox homeostasis and inhibited lipid accumulation in hepatocytes.ConclusionsShort-term exercise enhances BCAA catabolism in skeletal muscle and promotes glutamine production, which circulates to the liver to improve redox balance and alleviate MASH.

Targeting gut microbiota is an innovative approach to mitigate the development of metabolic dysfunction-associated steatotic liver disease-associated hepatocellular carcinoma (MASLD-HCC). This study aims to investigate the effects of Prohep, a probiotic mixture, both as a prophylactic measure and as an adjuvant therapy for low-dose sorafenib. A MASLD-HCC mice model was established by diethylnitrosamine (DEN) injection with feeding of a high-fat high-cholesterol (HFHC) diet. Gut microbiome profiles were later identified through shotgun sequencing. Our findings demonstrated that Prohep supplementation effectively suppressed MASLD-HCC development in mice. This protective effect was attributed to the modulation of gut microbiota and the increased production of short-chain fatty acids (SCFAs), propionate, and valerate. Prohep also activated AMPK, which decreased lipogenesis, reduced lipid uptake, and enhanced antioxidant enzyme expressions. Additionally, the cancer proliferation pathway PI3K/mTOR was inhibited in response to Prohep treatment. As an adjuvant therapy, Prohep improved the efficacy of low-dose sorafenib, as indicated by reduced tumor counts, alleviated inflammation, and increased hepatic superoxide dismutase (SOD) expression. The combination led to enhanced butyrate production, contributing to the overall therapeutic effects, thanks to the gut microbiota modulatory effects of Prohep. These results underscore Prohep's anti-tumorigenic properties and its potential to enhance the therapeutic outcomes of low-dose sorafenib in MASLD-HCC treatment. The study highlights the importance of gut microbiota modulation for developing effective neoadjuvant therapies and long-term management strategies for MASLD-HCC.

Abstract Metabolic dysfunction-associated steatotic liver disease related hepatocellular carcinoma (MASLD-HCC) is an emerging malignancy. However, the identity and function of cancer stem cells (CSCs) in MASLD-HCC are largely unclear. Here, we aim to investigate the function, mechanism, and targeted therapy of CSCs in MASLD-HCC. Intravital genetic lineage tracing in mice demonstrated that CD133+ cells from high-fat high cholesterol diet (HFHC)-induced MASLD-HCC possess the characteristics of CSCs, including superior self-renewal capacity in vitro and tumorigenicity in vivo compared to CD133- counterparts. ScRNA-seq of CD133+ progenies revealed their clonal expansion into multiple lineages during tumor progression, highlighting the role of CD133+ CSCs in contributing to cellular plasticity in MASLD-HCC. Further, we showed that CD133 functionally drives CSC phenotypes in MASLD-HCC and that hepatocyte-specific Cd133 overexpression in mice accelerated MASLD-HCC formation driven by diethylnitrosamine plus choline-deficient, high-fat (CDHF) diet. Mechanistically, CD133 interacts with myosin heavy chain 9 (MYH9) to promote the stabilization of active β-catenin, thereby propagating Wnt/β-catenin signaling critical for CSC phenotypes as well as pro-tumorigenic potential. Accordingly, MYH9 knockdown abrogated the induction of Wnt/β-catenin by CD133. In terms of translational relevance, targeted ablation of CD133+ cells using a selective diphtheria toxin receptor system (DTRlslCd133-CreER) in mice sensitizes MASLD-HCC to multikinase inhibitors (Sorafenib and Lenvatinib). Moreover, targeting of CD133 by nanoparticle-siRNA in combination with Sorafenib synergized to induce regression of tumors in subcutaneous and orthotopic MASLD-HCC. In summary, our results indicate that CD133+ CSCs potentiate MASLD-HCC by activating Wnt/β-catenin signaling through MYH9 and is a therapeutic target, especially in combination with multikinase inhibitors. Citation Format: Yasi Pan, Xiang Zhang, Chi Chun Wong, Huarong Chen, Kai Yuan, Jun Yu. Visualization and targeting of CD133+ cancer stem cells in MASLD-related hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_2):Abstract nr LB177.

Atherosclerosis (AS) is widely recognized as a consequence of chronic inflammation, with the nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome playing a pivotal role in mediating this inflammatory response. Kumquat flavonoids (KFs), the primary active ingredients in kumquat, have demonstrated potential in modulating inflammation and may help prevent AS. Herein, this study aimed to explore the protective effects and underlying mechanisms of KFs on AS using an ApoE-/- mouse model fed a high-fat/cholesterol diet (HFCD) and the mouse aortic vascular smooth muscle cell (MOVAS) inflammation model induced by oxidized low-density lipoprotein (ox-LDL). Our results show that KFs significantly reduced serum lipid levels and suppressed the overproduction of inflammatory cytokines in ApoE-/- mice. Notably, KFs also decreased the area of atherosclerotic lesions and plaque formation in the aorta of ApoE-/- mice. Additionally, in vivo (mouse aortic tissue) and in vitro (MOVAS cells), KFs were found to inhibit the activation of NLRP3 inflammasome and simultaneously upregulate microRNA-145 (miR-145). In conclusion, our findings suggest that KFs exert their inhibitory effects on NLRP3 inflammasome through upregulating miR-145, thereby alleviating the progression of AS.

Non-alcoholic fatty liver disease (NAFLD) is currently a public health problem that affects many people worldwide. New-generation dietary agents can contribute to disease control. Also, prediction of liver disease activity with serum markers is an important factor to reduce the need for liver biopsies. The aim of this study was to compare the effects of 20 mg/kg green coffee bean extract (GCBE, standardized to 10 mg/kg chlorogenic acid), 10 mg/kg chlorogenic acid (CA) and 50 mg/kg CA on NAFLD activity score (NAS). We also aimed to create a NAS prediction model as an alternative to liver biopsies. Male Wistar rats were fed either a high-fat high-cholesterol (HFHC) diet (NAFLD groups, n = 32) or a normal diet (control groups, n = 24). Green coffee bean extract (20 mg/kg) and CA (10 mg/kg and 50 mg/kg) were given by daily oral gavage. At the end of the study, blood samples and liver tissues were collected. Biochemical parameters, glyoxalase 1 (GLO1) and oxidative stress parameters were analyzed. An experienced histopathologist graded NAS. Green coffee bean extract and 50 mg/kg CA significantly reduced NAS (p < 0.01 and p < 0.001, respectively). HFHC-diet induced NAFLD was associated with higher serum GLO1 levels (p < 0.001). Serum malondialdehyde (MDA) levels were positively correlated with NAS while GLO1 was negatively correlated with NAS (p < 0.001, r = 0.698; p < 0.05, r = -0.367, respectively). Serum MDA and triglyceride were found to be statistically significant in predicting NAS (p < 0.001). Our study suggests that GCBE and CA can both ameliorate the development of NAFLD. Also, low-dose GCBE and high-dose CA showed similar hepatoprotective effects. Increased GLO1 in NAFLD may be a defense mechanism which is enhanced by GCBE and CA. Moreover, serum MDA and triglyceride levels are promising in NAS prediction.

LRP1 immunotherapy enhances cardiomyocyte respiration by restricting cholesteryl ester accumulation in mitochondria

Lifestyle interventions, such as diet and exercise, are currently the main therapies against metabolic dysfunction-associated steatotic liver disease (MASLD). However, not much is known about the combined impact of fiber and exercise on the modulation of gut-liver axis and MASLD amelioration. Here, we studied the impact of the combination of exercise training and a fiber-rich diet on the amelioration of MASLD. Male APOE*3-Leiden.CETP mice were fed a high-fat high-cholesterol diet with or without the addition of fiber (10% inulin) and exercise trained on a treadmill, or remained sedentary. Exercise training and fiber supplementation reduced fat mass gain and lowered plasma glucose levels. Only the combination treatment, however, induced fat loss and decreased plasma triglyceride and cholesterol levels compared with sedentary control mice. Exercise training with and without the addition of fiber had a similar ameliorating effect on the MASLD score. Only exercise without fiber decreased the hepatic expression of inflammatory markers. Fiber diet was mainly responsible for remodeling the gut microbial composition, with an increase in the relative abundance of the short-chain fatty acid (SCFA)-producing genera Anaerostipes and Muribaculaceae, whereas, surprisingly, exercise training alone and with fiber resulted in the highest increase of SCFA production. Overall, the combination of exercise training and dietary fiber decreases fat mass and improves glucose and lipid homeostasis but does not have an additional synergistic positive effect on liver health compared with exercise training alone.NEW & NOTEWORTHY The combination of dietary fiber intake and exercise training has a synergetic beneficial effect on the metabolic health, resulting in fat loss, lowered blood glucose, and lowered plasma lipid levels in mice with steatotic liver disease. However, fiber supplementation, despite a positive remodulation of the gut-liver axis, does not have an additional positive effect on liver health compared with exercise training alone.

ABSTRACT Obstructive sleep apnea (OSA) is characterized by intermittent hypoxia/hypercapnia (IHC), affects predominantly obese individuals, and increases atherosclerosis risk. Since we and others have implicated gut microbiota and metabolites in atherogenesis, we dissected their contributions to OSA-induced atherosclerosis. Atherosclerotic lesions were compared between conventionally-reared specific pathogen free (SPF) and germ-free (GF) Apoe−/− mice following a high fat high cholesterol diet (HFHC), with and without IHC conditions. The fecal microbiota and metabolome were profiled using 16S rRNA gene amplicon sequencing and untargeted tandem mass spectrometry (LC-MS/MS) respectively. Phenotypic data showed that HFHC significantly increased atherosclerosis as compared to regular chow (RC) in both aorta and pulmonary artery (PA) of SPF mice. IHC exacerbated lesions in addition to HFHC. Differential abundance analysis of gut microbiota identified an enrichment of Akkermansiaceae and a depletion of Muribaculaceae (formerly S24–7) family members in the HFHC-IHC group. LC-MS/MS showed a dysregulation of bile acid profiles with taurocholic acid, taurodeoxycholic acid, and 12-ketodeoxycholic acid enriched in the HFHC-IHC group, long-chain N-acyl amides, and phosphatidylcholines. Interestingly, GF Apoe−/− mice markedly reduced atherosclerotic formation relative to SPF Apoe−/− mice in the aorta under HFHC/IHC conditions. In contrast, microbial colonization did not show a significant impact on the atherosclerotic progression in PA. In summary, this research demonstrated that (1) IHC acts cooperatively with HFHC to induce atherosclerosis; (2) gut microbiota modulate atherogenesis, induced by HFHC/IHC, in the aorta not in PA; (3) different analytical methods suggest that a specific imbalance between Akkermansiaceae and Muribaculaceae bacterial families mediate OSA-induced atherosclerosis; and (4) derived bile acids, such as deoxycholic acid and lithocholic acid, regulate atherosclerosis in OSA. The knowledge obtained provides novel insights into the potential therapeutic approaches to prevent and treat OSA-induced atherosclerosis.