Atherosclerotic Plaque Formation Research Articles

Puerarin alleviates acrolein-induced atherosclerosis by activating the MYH9-mediated SIRT1/Nrf2 cascade to inhibit the activation of inflammasome.

Puerarin (Pue) has significant antioxidant and anti-inflammatory properties. This work was designed to clarify and investigate the potential mechanisms of Pue in atherosclerosis (AS) progression. In vivo, acrolein (Acr) was inhaled through drinking water to construct AS model. In vitro, CCK-8 assay and lactate dehydrogenase (LDH) assay kit were used to detect cell viability. Apoptosis was detected by flow cytometry. The content of malondialdehyde (MDA) was determined by commercial kit, the level of inflammatory factors was detected by ELISA, and proteins were determined by western blot. Pue administration could effectively reduce blood lipid level in Acr-fed mice. Pue suppressed oxidative stress, the formation of atherosclerotic plaques, and the process of aortic histological changes. Pue pretreatment decreased MDA in HUVECs and maintained the activity of antioxidant enzymes. Pue upregulated SIRT1/Nrf2 cascade in HUVECs. Pue increased MYH9 and inhibited NLRP3 inflammasome-related proteins, and the inhibition of MYH9 significantly impaired Pue-induced Nrf2 activation. Moreover, HUVEC cytotoxicity and apoptosis are alleviated by Pue, in addition to NLRP3-mediated pyroptosis in HUVECs induced by Acr. MYH9 inhibitors effectively suppressed the pyroptosis induced by Acr and prevented injury to HUVECs. In addition, Pue promoted SIRT1/Nrf2 cascade activation in HUVECs. Pue may alleviate Acr-induced AS by activating the MYH9-mediated SIRT1/Nrf2 cascade to inhibit inflammasome activation.

#302 Proteomic study reveals indoxyl sulfate induced changes in endothelial cells: role of extracellular vesicles in cardiovascular disease

Abstract Background and Aims Chronic kidney disease (CKD) lead to the accumulation of uremic toxins in the bloodstream, among which indoxyl sulfate (IS) is the most prominent. IS causes damage to endothelial cells lining blood vessels, ultimately leading to cardiovascular diseases in patients with CKD. Under physiological conditions, extracellular vesicles (EVs) are released by cells as intercellular communicators; however, after damage, the cargo of the EVs can be modified. This makes EV potential markers of damage, possibly carrying proteins of interest. This study aimed to investigate the impact of IS on endothelial cells and the release of EVs to identify potential therapeutic targets for preventing endothelial injury. Method Human umbilical vein endothelial cells (HUVEC) were cultured under control conditions and with IS exposure (250 μM IS, 24 h, n = 5). The released EVs were isolated from the supernatants by differential centrifugation and characterized by TEM and flow cytometry to confirm their identity as EVs (following the MISEV2018 guidelines). Proteome profiles of both cells and EVs were analyzed using a mass spectrometer (Orbitrap Exploris™ 240), followed by a Gene Set Enrichment Analysis (GSEA) which revealed altered functions. Results A total of 5871 proteins were identified in the proteomic analysis of cells, and 3614 proteins in EVs (with a False Discovery Rate (FDR) < 1%). In cells, 30 proteins were upregulated and 46 were found downregulated in response to IS, while in EVs, 51 proteins were found upregulated and 47 downregulated (IS vs C). GSEA revealed that IS potentiated adipogenesis (p-value < 0.001), TNF-α signaling via NFкB (p-value = 0.006), and xenobiotic metabolism in cells (p-value = 0.012). In contrast, EVs from cells treated with IS exhibited downregulation of extracellular matrix components (p-value = 0.003), myogenesis (p-value 0.009), genes downregulated in response to ultraviolet radiation (p-value = 0.014) and TNF-α signaling via NFкB (p-value = 0.041). Conclusion IS induces a change in protein expression in cells and their EVs, potentiating inflammation and atherosclerotic plaque formation with extracellular matrix loss. All these modifications contribute to endothelial dysfunction, which is the first step in the development of cardiovascular disease in patients with CKD. This study shed light on novel targets for intervention strategies to mitigate cardiovascular complications in patients with CKD. Funding Grants from the Instituto de Salud Carlos III (ISCIII) and cofounded by Fondos Europeo de Desarrollo Regional (FEDER): “PI19/00240” and “PI20/01321” and “FI20/00018” contract by the Instituto de Salud Carlos III and Ayuda de la Línea de Actuación “Excelencia para el Profesorado Universitario de la UAH” (EPU-INV-UAH/2022/001) from Alcala University.

Influence of polyphenolic compounds on human health and the course of a number of diseases

The purpose of this literature review is to elucidate the biological effects of polyphenols and their importance for human health. Polyphenols play the role of food antioxidants and are of growing interest in various fields of scientific research and including nutritional research due to the presence of these compounds in plant foods. In this review, a brief classification of polyphenols is presented. Polyphenols include a diverse group of compounds such as phenolic acids, flavonoids, stilbenes and lignans. Flavonoids are found in fruits such as apples, apricots, grapes and citrus fruits; vegetables are rich in phenolic acids. Polyphenols are also rich in nuts, legumes, green tea, blueberries, cocoa. Polyphenols are of interest because of their effective direct and indirect antioxidant mechanisms for attenuating chronic diseases, as well as leveling negative lifestyle and nutritional factors. The antioxidant properties of polyphenolic compounds limit oxidative stress, which is relevant in geriatrics. Through a diet enriched with polyphenols, it is possible to reduce the likelihood of developing cancer, and diabetes. The favorable effect of polyphenols is noted on the functioning of the cardiovascular system. Polyphenols protect the endothelium from damage, slow the formation of atherosclerotic plaques, exhibit antithrombotic action. Having antioxidant activity, polyphenols inhibit lipoperoxidation processes, having a therapeutic effect in Alzheimer's and Parkinson's disease. In particular, catechins are effective in Parkinson's disease and ferulic acid is effective in patients with Alzheimer's disease. The protective effects of polyphenols associated with their antioxidant activity make the use of these compounds in geriatrics attractive. Due not only to their antioxidant but also hypoglycemic properties, polyphenols may be an additional treatment for diabetes mellitus. The polyphenols theaflavins and thearubigins, found in black tea and possessing antioxidant properties, have anti-cancer effects. Polyphenols also protect the skin from sun damage. It can be concluded that patients with chronic diseases should be prescribed a diet rich in polyphenols.

The effect and mechanism of inulin on atherosclerosis is mediated by the characteristic intestinal flora and metabolites.

Inflammation and hyperlipidemia can cause atherosclerosis. Prebiotic inulin has been proven to effectively reduce inflammation and blood lipid levels. Utilizing a mouse model induced by a high-fat diet, this study aimed to explore whether the characteristic intestinal flora and its metabolites mediate the effects of inulin intervention on atherosclerosis and to clarify the specific mechanism. Thirty apolipoprotein E-deficient (ApoE-/-) mice were randomly divided into three groups. They were fed with a normal diet, a high-fat diet or an inulin+high-fat diet for 16 weeks. The total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) in the three groups were compared. The gross aorta and aortic sinus of mice were stained with oil red O, and the area of atherosclerotic plaque was observed and compared. The diversity and structure of the mouse fecal flora were detected by sequencing the V3-V4 region of the 16S rRNA gene, and the levels of metabolites in mouse feces were assessed by gas chromatography-mass spectrometry. The plasma lipopolysaccharide (LPS) levels and aortic inflammatory factors were measured by multi-index flow cytometry (CBA). ApoE-/- mice fed with the high-fat diet exhibited an increase of approximately 46% in the area of atherosclerotic lesions, and the levels of TC, TG and LDL-C were significantly increased (P < 0.05) compared with levels in the normal diet group. After inulin was added to the high-fat group, the area of atherosclerotic lesions, the level of serum LPS and aortic inflammation were reduced, and the levels of TC, TG and LDL-C were decreased (P < 0.05). Based on 16S rRNA gene detection, we found that the composition of the intestinal microbiota, such as Prevotella, and metabolites, such as L-arginine, changed significantly due to hyperlipidemia, and the dietary inulin intervention partially reversed the relevant changes. Inulin can inhibit the formation of atherosclerotic plaques, which may be related to the changes in lipid metabolism, the composition of the intestinal microbial community and its metabolites, and the inhibition of the expression of related inflammatory factors. Our study identified the relationships among the characteristic intestinal microbiota, metabolites and atherosclerosis, aiming to provide a new direction for future research to delay or treat atherosclerosis by changing the composition and function of the host intestinal microbiota and metabolites.

LOX-1 in Cardiovascular Disease: A Comprehensive Molecular and Clinical Review.

LOX-1, ORL-1, or lectin-like oxidized low-density lipoprotein receptor 1 is a transmembrane glycoprotein that binds and internalizes ox-LDL in foam cells. LOX-1 is the main receptor for oxidized low-density lipoproteins (ox-LDL). The LDL comes from food intake and circulates through the bloodstream. LOX-1 belongs to scavenger receptors (SR), which are associated with various cardiovascular diseases. The most important and severe of these is the formation of atherosclerotic plaques in the intimal layer of the endothelium. These plaques can evolve into complicated thrombi with the participation of fibroblasts, activated platelets, apoptotic muscle cells, and macrophages transformed into foam cells. This process causes changes in vascular endothelial homeostasis, leading to partial or total obstruction in the lumen of blood vessels. This obstruction can result in oxygen deprivation to the heart. Recently, LOX-1 has been involved in other pathologies, such as obesity and diabetes mellitus. However, the development of atherosclerosis has been the most relevant due to its relationship with cerebrovascular accidents and heart attacks. In this review, we will summarize findings related to the physiologic and pathophysiological processes of LOX-1 to support the detection, diagnosis, and prevention of those diseases.

Broader Perspective on Atherosclerosis-Selected Risk Factors, Biomarkers, and Therapeutic Approach.

Atherosclerotic cardiovascular disease (ASCVD) stands as the leading cause of mortality worldwide. At its core lies a progressive process of atherosclerosis, influenced by multiple factors. Among them, lifestyle-related factors are highlighted, with inadequate diet being one of the foremost, alongside factors such as cigarette smoking, low physical activity, and sleep deprivation. Another substantial group of risk factors comprises comorbidities. Amongst others, conditions such as hypertension, diabetes mellitus (DM), chronic kidney disease (CKD), or familial hypercholesterolemia (FH) are included here. Extremely significant in the context of halting progression is counteracting the mentioned risk factors, including through treatment of the underlying disease. What is more, in recent years, there has been increasing attention paid to perceiving atherosclerosis as an inflammation-related disease. Consequently, efforts are directed towards exploring new anti-inflammatory medications to limit ASCVD progression. Simultaneously, research is underway to identify biomarkers capable of providing insights into the ongoing process of atherosclerotic plaque formation. The aim of this study is to provide a broader perspective on ASCVD, particularly focusing on its characteristics, traditional and novel treatment methods, and biomarkers that can facilitate its early detection.

Exploring the role of myeloperoxidase in the atherosclerotic process in hypoxic mice based on the MAPK signaling pathway

Atherosclerosis (AS) is the common pathophysiological basis of various cardiovascular diseases and the leading cause of death from cardiovascular disease worldwide. When the body is in a hypoxic environment, enhanced oxidative stress and significant accumulation of reactive oxygen species (ROS) in tissue cells exacerbate the inflammatory response, resulting in increased release of myeloperoxidase (MPO), catalyzing the formation of large quantities of hypochlorous acid (HOCl), further oxidative modification of low-density lipoprotein (LDL), and exacerbating the formation and progression of atherosclerotic plaques. The MAPK signaling pathway is important in oxidative stress-mediated promotion of atherogenesis. MPO -/- mice were used in this study to establish a hypoxia model simulating 5000 m altitude and a Western high-fat diet-induced atherosclerosis model for 12 weeks. Exploring the role of MPO in the atherosclerotic process in hypoxic mice by observing the MAPK signaling pathway to provide a therapeutic target for the prevention and treatment of hypoxic atherosclerotic disease in the plateau. We found that hypoxia promotes the formation of atherosclerosis in mice, and the mechanism may be that increased MPO in vivo promotes an inflammatory response, which plays a crucial role in the formation of atherosclerosis. In addition, hypoxia further exacerbates plaque instability by activating the MAPK signaling pathway to upregulate vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP9), which in turn promotes angiogenesis within the plaque. Therefore, a potential target for preventing and treating hypoxic atherosclerotic disease is the inhibition of MPO.

The role of γδT lymphocytes in atherosclerosis.

Atherosclerosis poses a significant threat to human health, impacting overall well-being and imposing substantial financial burdens. Current treatment strategies mainly focus on managing low-density lipids (LDL) and optimizing liver functions. However, it's crucial to recognize that Atherosclerosis involves more than just lipid accumulation; it entails a complex interplay of immune responses. Research highlights the pivotal role of lipid-laden macrophages in the formation of atherosclerotic plaques. These macrophages attract lymphocytes like CD4 and CD8 to the inflamed site, potentially intensifying the inflammatory response. γδ T lymphocytes, with their diverse functions in innate and adaptive immune responses, pathogen defense, antigen presentation, and inflammation regulation, have been implicated in the early stages of Atherosclerosis. However, our understanding of the roles of γδ T cells in Atherosclerosis remains limited. This mini-review aims to shed light on the characteristics and functions of γδ T cells in Atherosclerosis. By gaining insights into the roles of γδ T cells, we may uncover a promising strategy to mitigate plaque buildup and dampen the inflammatory response, thereby opening new avenues for effectively managing this condition.

Macrophage-Derived GSDMD Plays an Essential Role in Atherosclerosis and Cross Talk Between Macrophages via the Mitochondria-STING-IRF3/NF-κB Axis.

Macrophages play a crucial role in atherosclerotic plaque formation, and the death of macrophages is a vital factor in determining the fate of atherosclerosis. GSDMD (gasdermin D)-mediated pyroptosis is a programmed cell death, characterized by membrane pore formation and inflammatory factor release. ApoE-/- and Gsdmd-/- ApoE-/- mice, bone marrow transplantation, and AAV (adeno-associated virus serotype 9)-F4/80-shGSDMD (shRNA-GSDMD) were used to examine the effect of macrophage-derived GSDMD on atherosclerosis. Single-cell RNA sequencing was used to investigate the changing profile of different cellular components and the cellular localization of GSDMD during atherosclerosis. First, we found that GSDMD is activated in human and mouse atherosclerotic plaques and Gsdmd-/- attenuates the atherosclerotic lesion area in high-fat diet-fed ApoE-/- mice. We performed single-cell RNA sequencing of ApoE-/- and Gsdmd-/- ApoE-/- mouse aortas and showed that GSDMD is principally expressed in atherosclerotic macrophages. Using bone marrow transplantation and AAV-F4/80-shGSDMD, we identified the potential role of macrophage-derived GSDMD in aortic pyroptosis and atherosclerotic injuries in vivo. Mechanistically, GSDMD contributes to mitochondrial perforation and mitochondrial DNA leakage and subsequently activates the STING (stimulator of interferon gene)-IRF3 (interferon regulatory factor 3)/NF-κB (nuclear factor kappa B) axis. Meanwhile, GSDMD regulates the STING pathway activation and macrophage migration via cytokine secretion. Inhibition of GSDMD with GSDMD-specific inhibitor GI-Y1 (GSDMD inhibitor Y1) can effectively alleviate the progression of atherosclerosis. Our study has provided a novel macrophage-derived GSDMD mechanism in the promotion of atherosclerosis and demonstrated that GSDMD can be a potential therapeutic target for atherosclerosis.

Abstract 1039: Endothelial Deletion Of Nck1 Reduces Atherosclerosis In Apoe-/- Mice Through The Nck1-atf3 Axis

The Nck family of signaling adaptors, including both Nck1 and Nck2, classically regulate cell motility during vascular development and postnatal angiogenesis. While Nck1 and Nck2 play redundant roles in this regard with deletion of both isoforms required to affect tissue remodeling, we demonstrated that global Nck1 deletion was sufficient to reduce atherosclerotic plaque formation and selective Nck1 inhibition reduces proinflammatory endothelial activation in response to oscillatory shear stress. Therefore, we sought to determine the role of endothelial Nck1 in atherosclerosis and ischemic angiogenesis using novel endothelial-specific Nck1 knockout mice. Endothelial-specific Nck1 deletion significantly reduced Western diet-induced atherosclerotic plaque formation at multiple sites compared to mice expressing endothelial-specific Cre alone or to endothelial-specific Nck2 knockout mice. This reduced plaque formation was characterized by diminished macrophage and smooth muscle incorporation despite enhanced weight gain in endothelial Nck1 knockout mice. RNA Sequencing analysis of endothelial cells in culture showed that Nck1 depletion reduces the expression of a variety of proinflammatory pathways but enhances the expression of genes involved in central carbon metabolism and proliferation. While Nck1 deletion is sufficient to blunt atherogenic inflammation, Nck1 inhibition did not affect angiogenesis or limb perfusion in the femoral artery ligation model of hind limb ischemia. To further characterized Nck1’s proinflammatory role, we utilized affinity precipitation-mass spectrometry and identified Activating Transcription Factor 3 ATF3 as a Nck1-selective binding partner. While no studies to date have examined endothelial ATF3 signaling in atherosclerosis, our preliminary data show that Nck1 is required for ATF3 expression in response to disturbed flow in vitro and in vivo , but the mechanism by which disturbed flow stimulates ATF3 expression remains unknown. ATF3 knockdown reduces disturbed flow-induced proinflammatory gene expression, suggesting that the Nck1-ATF3 signaling axis is a novel regulator of flow-induced inflammation.

Abstract 2069: Impact Of Endothelial Stiffening And CD36-mediated Lipid Uptake On Atherosclerotic Lesion Formation

Objective: To determine the impact of endothelial stiffening induced by CD36 (cluster of differentiation 36) lipid uptake in the development of atherosclerosis. Approach and Results: Endothelial stiffening of intact en face monolayers, as assessed by atomic force microscopy, reveal the hyperlipidemic mouse model of atherosclerosis, LDLR -/- (low density lipoprotein receptor knockout) is significantly stiffer, especially in the atheroprone aortic arch, as compared to wild type controls (n=4 male mice with 8-15 tissue measurement sites per condition per mouse, p&lt;0.05). Furthermore, a 4-5 month high fat diet (HFD) feeding in LDLR -/- mice resulted in significantly enhanced stiffening compared to regular chow feeding, an effect that was observed in males but not females (n=6-8 mice, p&lt;0.05). There was a significantly greater plaque development, as analyzed using Oil O Red staining for lipids, in male HFD-fed LDLR -/- mice, as compared to age-matched females (n=5-6, p&lt;0.05). No significant difference between males and females was found in lipid panel parameters (n=4-6) or weight gain (n=10-14). Specific downregulation of CD36 in male hypercholesterolemic Cdh5.CreERT2CD36fl/fl LDLR -/- mice led to both a significant decrease in endothelial stiffening (n=5 mice, p&lt;0.05) and in the areas of atherosclerotic lesion formation (n=5-6 mice, p&lt;0.05). Mechanistically, we show that the saturated fatty acid, palmitic acid, increased endothelial stiffening in the en face monolayers of wild type mice, an effect which is prevented by the pre-treatment of the CD36 irreversible inhibitor, SSO (sulfo-N-succinimidyl oleate, n=5-6 male mice, p&lt;0.05). Conclusions: These results highlight that endothelial stiffness from intact aortas and atherosclerotic plaque formation following HFD feeding is strongly dependent on endothelial CD36.

Abstract 1053: The Role Of Bile Acid-initiated Signaling In Vascular Smooth Muscle Cell Phenotypic Switching And Atherosclerosis

Atherosclerosis is a chronic inflammatory disease in which foam cells (lipid-laden macrophage-like cells) accumulate in the arterial wall. Vascular smooth muscle cells (VSMCs) are not terminally differentiated; they switch to a foam cell-like phenotype in response to various stimuli, such as cholesterol (CHOL), increasing susceptibility to vascular diseases. Modulated SMCs are the main contributors to atherosclerotic plaque formation. The intracellular metabolism of CHOL via peroxisomes in hepatocytes produces bile acids like cholic or deoxycholic acid, which agonize liver X receptor (LXR). LXR facilitates the first step of reverse CHOL transport. It is unknown if VSMCs synthesize bile acids and whether peroxisomes and the bile acids they produce play a role in VSMC-derived foam cell formation. Using human aortic SMCs (HASMCs), we found that CHOL treatment decreased expression of several peroxisomal proteins. Peroxisomal loss-of-function experiments demonstrated increased intracellular lipid accumulation, as visualized with Oil Red O (ORO) staining. We have shown that HASMCs synthesize bile acids after exposure to CHOL, which led us to hypothesize that bile acid-initiated signaling participates in the phenotypic modulation initiated by CHOL in VSMCs. Treatment of HASMCs with LXR agonists or bile acids increased expression of the efflux CHOL transporter ABCA1, bile acid-related peroxisomal enzymes, and differentiation markers; they decreased expression of macrophage and foam cell markers and ORO staining. These data show that VSMCs have the metabolic capacity to synthesize bile acids, which is impaired if peroxisomes are non-functional or absent. Our data indicates that activation of bile acid receptors upregulates ABCA1, favoring a differentiated VSMC phenotype. Together, these data suggest that activation of LXR plays a role in CHOL-induced VSMC to foam cell transition and that LXR is a potential therapeutic target which promotes VSMC differentiation and decreases atherosclerotic plaque formation.

Abstract 3078: Free Fatty Acid Receptor 4: An Emerging Target To Reduce Atherosclerosis

Free fatty acid receptor 4 (FFAR4) has recently emerged as a target for preventing CVD through its ability to reduce macrophage foam cell formation and inflammation. We have shown that activation of FFAR4 by synthetic agonists decreases foam cell formation, whereas deficiency of FFAR4 increases foam cell formation. Building on these data, we hypothesized that FFAR4 would protect against atherosclerotic plaque formation in vivo . To test this hypothesis, we generated global FFAR4 knockout mice on an apoE-deficient background. Male and female apoE/FFAR4 double knockout mice (n=20) and their apoE knockout controls (n=20) were fed a Western diet (42% calories from fat and 0.2% cholesterol) for 16 weeks. We collected weekly body weights and fasting plasma samples from mice at 0, 8, and 16 weeks to measure triglyceride and cholesterol levels. At the end of the 16-week study, atherosclerotic plaque severity was determined by analyzing the aortic sinus lesion area of the heart and the en-face lesion area of the aortic arch. In both the male and female mice, we observed no differences in weekly body weights or lipid levels between the apoE knockout and apoE/FFAR4 double knockout mice. In our first cohort of male mice (n=10), the two genotypes showed no difference in the aortic sinus lesion area. However, male apoE/FFAR4 double knockout mice had a statistically significant (p=0.039) 1.4-fold increase in total aortic arch lesion area compared to FFAR4-deficient mice. While female mice (n=9) showed no differences in atherosclerotic plaque in the aortic sinus and aortic arch between the two genotypes, they did present with 1.7-fold (p=0.002) and 2-fold increases (p=0.009) in aortic sinus lesion area and aortic arch lesion area, respectively, as compared to male mice. Based on analysis of an initial set of mice, our data support a protective role for FFAR4 in the development of atherosclerotic plaque formation in male mice, and our findings may have uncovered a new therapeutic target against atherosclerosis.

Abstract 3019: Piezo-1 Expression Decreases Under Disturbed Flow Model Of Femoral Atherosclerosis.

Background: Peripheral arterial disease (PAD) is often manifest by focal atherosclerotic plaque formation in the infra-inguinal arteries. However, murine models of atherosclerosis in the infra-inguinal arteries are lacking. It is well known that low and oscillatory wall shear stress, which occurs around blockages in PAD, promotes focal atherosclerotic plaque, but this has not been tested in vivo. Piezo1 is a mechanosensitive ion channel associated with hemodynamic changes in the arterial wall, but its role in flow-mediated atherosclerotic plaque deposition is not known. Objective: To develop a flow-mediated murine model of focal atherosclerotic plaque in the femoral arteries. Methods: 8 male and female S129 mice underwent partial femoral artery ligation (Figure A/B). Wall shear stress was calculated from duplex ultrasound imaging (Figure C/D). Atherogenic conditions were imposed with PCSK9 infection and high fat diet; this was initiated 1 week prior to partial femoral ligation. Histologic comparison of the left (partial femoral ligation) and right (normal stable flow) was performed. Results: Low wall shear stress was induced in this model (Figure C/D). Piezo1 expression was decreased in both male and female arteries under these conditions in this model (Figure E). Atherosclerotic plaque was induced by the partial femoral artery ligation but not on the normal stable flow side (Figure F). Conclusions: We have successfully developed a flow-mediated model of focal atherosclerotic plaque in murine femoral arteries. This is associated with downregulation of Piezo1 gene expression. Ongoing work is examining the causative role of Piezo1 on endothelial response to disturbed flow.

PNPLA3 I148M Promotes Fatty Liver Disease While Preserving Cardiac Health

Metabolic-associated fatty liver disease (MAFLD) and cardiovascular disease (CVD) are commonly associated comorbidities. A common genetic variant in the PNPLA3 gene, rs738409 (I148M), is the greatest genetic risk factor for development and progression of MAFLD; however, MAFLD patients expressing PNPLA3 I148M display modest protection against coronary artery disease (CAD) and adverse cardiovascular events relative to wildtype carriers. Currently, the exact mechanisms by which PNPLA3 I148M causes MAFLD and preserves cardiac health are not well understood. Previous work suggests that the variant increases susceptibility to fatty liver disease by altering hepatic lipid metabolism, which may explain a potential mechanism for its cardioprotective effects. In order to investigate how PNPLA3 I148M might increase susceptibility to MAFLD while reducing risk of CVD we expressed either human wildtype (WT) PNPLA3 or PNPLA3 I148M as well as a GFP control specifically in the livers of PNPLA3 knockout mice. Our data demonstrate that hepatic expression of PNPLA3 I148M reduces secretion of VLDL TAGs from adipose tissue derived fatty acids. In agreement with prior investigations, mice expressing PNPLA3 I148M and fed a chow diet demonstrated significant increases in hepatic accumulation of neutral lipids (triglycerides &amp; cholesterol esters) relative to WT and GFP expressing mice. Similarly, when fed a fatty liver promoting diet (D009100310) high in fructose, cholesterol, and saturated fats there remained a strong trend for greater liver to body weight ratios and hepatic TAGs in mice expressing the variant when compared to WT expressing mice (p=0.052 and p=0.071 respectively). Changes in hepatic lipid accumulation were not due to any alterations in insulin sensitivity, daily food consumption, energy expenditure or oxygen consumption between WT and I148M groups. Echocardiography revealed that hearts of mice expressing WT PNPLA3 had increases in left ventricular thickness (p=0.051) and left ventricle weights following dietary treatment that was not apparent in the I148M expressing mice. Moreover, there was a reduction in E/A ratios, a sensitive measure of diastolic functioning, in GFP and WT expressing mice post-treatment, that was not seen in the I148M mice. Overall, these findings indicate that PNPLA3 I148M promotes fatty liver, yet protects against diet-induced effects on ventricular remodeling and diastolic functioning. Our next steps will be to assess measures of atherosclerotic plaque formation around major blood vessels and investigate the metabolic pathways altered by PNPLA3 I148M. NIH R00 DK114471 NIH R01 DK126743 USDA grant no. 2021-09425 Pilot feasibility grant from University of Michigan: P30DK020572 Detroit Cardiovascular Training grant NIH 2T32HL120822. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Proteomic Signatures of Coronary Artery Disease and Hypercholesterolemia

Background: Coronary artery disease (CAD) is the most common type of cardiovascular disease (CVD), it causes narrowing of the blood vessel lumen due to atherosclerotic plaque formation. Atherosclerosis, is strongly influenced by hypercholesterolemia (HC). Proteomics offers advantages over traditional studies that focus on individual biomarkers. This method provides a holistic view by examining the entire proteomic spectrum in a biological sample, making it an indispensable asset for spotting potential novel protein biomarkers directly linked to atherosclerosis occurrence, progression, and complications such as plaque instability or rupture. Methods: Slow Off-rate Modified Aptamer (SOMAmer)-based protein array was used to quantify proteins. Our cross-sectional study involved healthy controls (n=45), and patients diagnosed with HC (n=51) or CAD (n=32). Proteome data was analyzed using multiple statistical methods. Results: The orthogonal partial least square discriminant analysis (OPLS-DA) revealed a clear separation of the 3 study groups based on two principal components, with R2X= 0.267, R2Y= 0.9, and Q2= 0.28, indicating distinct protein alterations between groups. A total of 1,305 proteins were analyzed and 156 of them in CAD vs control, 140 of them in HC vs control, and 326 of the proteins in CAD vs HC comparisons, were significantly differentially expressed with a p-value&lt;0.05. Among those, q-value&lt;0.05 were determined. As a result, in CAD patients, 2 proteins exhibited significantly differential expressions compared to healthy controls. In contrast, HC patients had 15 significantly altered proteins compared to controls. Furthermore, a comparison between CAD and HC patients revealed 87 significantly regulated proteins. The receiver operating characteristic (ROC) was performed on proteins with distinct expressions ( q&lt;0.05) in pairwise group comparisons. The AUC was calculated to identify potential biomarker candidates for the disease groups. In the CAD vs. control analysis, both significant proteins (PCSK9, SDF-1) exhibited an AUC of 0.75. In the comparison between HC and control groups, out of 15 significantly altered proteins, 7 had an AUC of 0.75 or higher (LRP1B, Apo E, QORL1, Apo E3, Apo B, Transferrin, MMP-3). Among the 87 proteins significantly differentially expressed in CAD versus HC, 59 had an AUC ≥ 0.75. Sixteen of these proteins had an AUC ≥ 0.80. Conclusion: Distinct protein patterns associated with each condition were identified. CAD patients showed a significant increase in the cholesterol-metabolizing protein PCSK9 and varying levels of the angiogenesis-related protein SDF-1. Several other proteins (LRP1B, VAV, CSRP3, HSP 60, and TAK1-TAB1) also exhibited unique expressions in disease groups, suggesting potential roles in atherosclerosis development. Moreover, a statistically significant correlation between SDF-1, ApoB levels, and LDL levels was found in CAD patients but not in healthy controls. This discovery phase cohort study unveils potential biomarkers for atherosclerosis within the HC context. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

ASYMPTOTIC STABILITY FOR A FREE BOUNDARY MODEL OF AN ATHEROSCLEROTIC PLAQUE FORMATION IN THE PRESENCE OF REVERSE CHOLESTEROL TRANSPORT WITH HOLLING TYPE-III FUNCTIONAL RESPONSE

Atherosclerosis, as a chronic inflammatory disease, has been a threat to human health. How to diagnose and prevent this disease has long been the focus of medical and biomathematics study. In this paper, we investigate a free boundary model of an atherosclerotic plaque formation in the presence of reverse cholesterol transport, which includes low-density lipoprotein, high-density lipoprotein, endothelial stimulating cytokines, pro-inflammatory and anti-inflammatory macrophages, as well as foam cells. For this model, we use the Holling type-III response function instead of the usual Holling type-II to describe the change of the concentration of each substance. We first introduce the auxiliary function [Formula: see text] to prove the existence and uniqueness of small radially symmetric stationary plaque for appropriate [Formula: see text] and [Formula: see text] by using the contraction mapping principle and maximum principle, and then establish a condition to ensure their asymptotic stability behavior by expanding the corresponding steady-state solution.

Engineering Nanoplatforms for Theranostics of Atherosclerotic Plaques.

Atherosclerotic plaque formation is considered the primary pathological mechanism underlying atherosclerotic cardiovascular diseases, leading to severe cardiovascular events such as stroke, acute coronary syndromes, and even sudden cardiac death. Early detection and timely intervention of plaques are challenging due to the lack of typical symptoms in the initial stages. Therefore, precise early detection and intervention play a crucial role in risk stratification of atherosclerotic plaques and achieving favorable post-interventional outcomes. The continuously advancing nanoplatforms have demonstrated numerous advantages including high signal-to-noise ratio, enhanced bioavailability, and specific targeting capabilities for imaging agents and therapeutic drugs, enabling effective visualization and management of atherosclerotic plaques. Motivated by these superior properties, various noninvasive imaging modalities for early recognition of plaques in the preliminary stage of atherosclerosis are comprehensively summarized. Additionally, several therapeutic strategies are proposed to enhance the efficacy of treating atherosclerotic plaques. Finally, existing challenges and promising prospects for accelerating clinical translation of nanoplatform-based molecular imaging and therapy for atherosclerotic plaques are discussed. In conclusion, this review provides an insightful perspective on the diagnosis and therapy of atherosclerotic plaques.

The Role of Macrophage Dynamics in Atherosclerosis Analyzed Using a Petri Net-Based Model

Atherosclerosis, a chronic inflammatory and oxidative stress-mediated disease impacting the arterial system, stands as a primary cause of morbidity and mortality worldwide. The complexity of this disease, driven by numerous factors, requires a thorough investigation of its underlying mechanisms. In our study, we explore the complex interplay between cholesterol homeostasis, macrophage dynamics, and atherosclerosis development using a Petri net-based model anchored in credible, peer-reviewed biological and medical research. Our findings underscore the significant role of macrophage colony-stimulating factor (M-CSF) inhibition in reducing atherosclerotic plaque formation by modulating inflammatory responses and lipid accumulation. Furthermore, our model highlights the therapeutic potential of targeting the C-X-C motif ligand 12 (CXCL12)/ C-X-C motif chemokine receptor type 4 (CXCR4) pathway to hinder hematopoietic stem and progenitor cells’ (HSPCs’) mobilization and plaque development. Based on the results obtained, which are in agreement with current studies, additional strategies are also proposed, such as decreasing M1 macrophage polarization for therapeutic gains, opening the door to future research and novel treatment approaches.

Gut microbiome and metabolomic profiles reveal the antiatherosclerotic effect of indole-3-carbinol in high-choline-fed ApoE-/- mice

BackgroundThe metabolites produced from choline contribute to atherosclerosis (AS) pathogenesis, and the gut microbiota is redundantly essential for this process. Indole-3-carbinol (I3C), found in cruciferous vegetables such as broccoli, cabbage, cauliflower and brussels sprouts, helps prevent hyperlipidemia, maintain the gut microbiota balance, and decrease the production of trimethylamine-N-oxide (TMAO) from choline in the diet. PurposeThe objective of this research was to investigate the impact of I3C on choline-induced AS and to further elucidate the underlying mechanism involved. MethodsAS models of high-choline-induced ApoE-/- mice and TMAO-promoted foamy macrophages were established to observe the effect of I3C on the formation of atherosclerotic plaques and foam cells and changes in AS-related indicators (including blood biochemical indicators, TMA, TMAO, SRA, and SRB1), and integrated analyses of the microbiome and metabolome were used to reveal the mechanism of action of I3C. ResultsWe found that I3C inhibited high-choline-induced atheroma formation (50–100 mg/kg/d, in vivo) and slightly improved the lipid profile (15 mg/kg/d, in vivo). Moreover, I3C suppressed lipid influx at a concentration of 40 µmol/L in vitro, enhanced the diversity of the gut microbiota and the abundance of the phylum Verrucomicrobia, and consequently modified the gut microbial metabolites at a dosage of 50 mg/kg/d in the mice. Associative analyses based on microbiome and metabolomics revealed that 1-methyladenosine was a key modulator of the protective effect of I3C against AS in high-choline-induced ApoE-/- mice. ConclusionThese findings demonstrate for the first time that I3C ameliorates AS progression through remodeling of the gut microbiome and metabolomics, which paves the way for the possible therapeutic use of this vegetable-derived natural compound and may reduce the clinical severity of AS-related cardiovascular diseases.