Atherosclerotic Plaque Formation Research Articles

Abstract 705: Epha2 Ligand-dependent And Ligand-independent Signaling In Vascular Smooth Muscle Cell Proliferation And Migration

During atherosclerotic plaque formation, vascular smooth muscle cells (VSMCs) undergo phenotypic modulation from a contractile, quiescent phenotype to a proliferative and synthetic phenotype, allowing VSMCs to deposit form a protective fibrous cap to stabilize the cap through extracellular matrix assembly. Our lab discovered a novel role for the receptor tyrosine kinase EphA2 in SMC phenotypic modulation and proliferation both in vitro and in vivo . In many cancer models, EphA2 can act as a tumor promoter in the absence of ligand and a tumor suppressor in the presence of its ephrinA1 ligand. Therefore, we sought to determine the role of ligand-dependent and ligand-independent signaling in VSMC function. While EphA2 expression is required for maximal VSMC proliferation and migration, EphA2 ligation with ephrinA1 inhibits VSMC proliferation (associated with diminished Erk1/2 and Akt activation) and migration. While ligand treatment induces EphA2 tyrosine phosphorylation, treating VSMCs with serum promotes robust S897 phosphorylation (marker for ligand-independent signaling) specifically through RSK2, whereas no role was found for other described S897 kinases (e.g. Akt, RSK1). EphA2 Ser897 phosphorylation specifically localizes to lamellipodia in VSMCs treated with serum, and blocking RSK2-dependent S897 phosphorylation significantly inhibited VSMC migration but did not affect VSMC proliferation. Taken together, these data suggest that VSMC EphA2 ligation suppresses proliferation and migration, whereas RSK2-dependent EphA2 Ser897 phosphorylation in the absence of ligand promotes VSMC migration, and EphA2 expression likely enhances VSMC proliferation through a ligand-independent response independent of S897 phosphorylation.

Abstract 146: Foamy Macrophages Rely On Trem2 For Survival During Atherosclerosis

Atherosclerotic plaque formation is supported by the accumulation of cholesterol loaded foamy macrophages within the artery wall. Molecular regulators of foamy macrophage differentiation and survival remain to be fully understood. We performed trajectory analysis of a murine META scRNAseq dataset and compared it against a genome-wide foamy macrophage CRISPR screen, which identified Triggering Receptor Expressed on Myeloid Cells 2 (Trem2) as a candidate regulator for foamy macrophage lipid uptake and differentiation. Conditional deletion of Trem2 in macrophages led to dramatically reduced atherosclerotic plaque burden in the aorta and aortic root after 8 or 16 weeks of western diet feeding. This was associated with reduced local macrophage proliferation and enhanced cell death within lesions, but independent of circulating cholesterol or inflammatory markers. In addition, Trem2-deficient macrophages showed reduced ability to take up lipids compared to WT macrophages, both in vivo and in vitro. Gene expression analysis of foamy and non-foamy macrophages revealed that Trem2 was required for proliferation gene expression and for the induction of cholesterol efflux genes after lipid loading. Without Trem2 foamy macrophages accumulated excess lipid in the endoplasmic reticulum (ER), leading to an ER-stress response and cholesterol-mediated cytotoxicity. Overall, this study reveals Trem2 as a mediator of foamy macrophage differentiation and atherosclerosis plaque growth, and furthermore identifies the Trem2 pathway as a putative therapeutic target for future intervention studies.

Abstract 710: Specific Removal Of Oxidized Cholesterol With A Cyclodextrin Dimer Drug Candidate Towards Reversing Atherosclerosis

Cardiovascular disease (CVD) is the world’s biggest killer. Current treatments for CVD are failing to deliver on promises to beat this deadly and complex family of diseases. New approaches that go beyond targeting LDL are needed. Cyclarity Therapeutics has developed a novel class of specifically engineered, dimerized cyclodextrin (CD) molecules for the encapsulation of toxic oxidized cholesterol. Oxidized cholesterol accumulates over time and causes dysfunction in many cell types, linking it to several age-related diseases including atherosclerosis. Presently, treatments for atherosclerosis are invasive, expensive, and/or show limited benefits. Here, a combination of in silico, in vitro, and ex vivo methods are used to implement a synergistic rational drug design strategy for developing CDs to remove atherogenic oxidized cholesterol (primarily 7-ketocholesterol (7KC)) from cells and tissues. Our results thus far indicate that our lead compound, UDP-003, can both prevent and reverse the formation of atherogenic foam cells in-vitro. UDP-003 binds and clears 7KC from in-vivo and ex-vivo systems selectivity. IND enabling studies, including pivotal GLP studies in rodent and non-rodent species, are complete and show that UDP-003 has an excellent safety profile with no significant clinical liabilities. UDP-003 has been awarded the Innovative Licensing and Access Pathway (ILAP) Innovation Passport by the MHRA, which is accelerating Cyclarity’s path to clinic and will facilitate rapid and innovative clinical trials. In totality, our data suggest that targeted removal of 7KC from foam cells with our proprietary class of cyclodextrin compounds has the potential to prevent and reverse the formation of atherosclerotic plaques. This innovation represents the first disease-modifying therapeutic approach to treating atherosclerotic disease.

Predicting coronary atherosclerosis heart disease with pericoronary adipose tissue attenuation parameters based on dual-layer spectral detector computed tomography: a preliminary exploration.

Coronary atherosclerosis is a chronic inflammatory condition. Pericoronary adipose tissue (PCAT) attenuation is closely related to coronary inflammation. This study aimed to investigate the relationship between PCAT attenuation parameters and coronary atherosclerotic heart disease (CAD) using dual-layer spectral detector computed tomography (SDCT). This cross-sectional study included eligible patients who underwent coronary computed tomography angiography using SDCT at the First Affiliated Hospital of Harbin Medical University between April 2021 and September 2021. Patients were classified as CAD (with coronary artery atherosclerotic plaque) or non-CAD (without coronary artery atherosclerotic plaque). Propensity score matching was used to match the two groups. The fat attenuation index (FAI) was used to quantify PCAT attenuation. The FAI was measured on conventional images (120 kVp) and virtual monoenergetic images (VMI) by semiautomatic software. The slope of the spectral attenuation curve (λ) was calculated. Regression models were established to evaluate the predictive value of PCAT attenuation parameters for CAD. A total of 45 patients with CAD and 45 patients without CAD were enrolled. The PCAT attenuation parameters in the CAD group were significantly higher than those in the non-CAD group (all P values <0.05). The PCAT attenuation parameters of vessels with or without plaques in the CAD group were higher than those of vessels without plaques in the non-CAD group (all P values <0.05). In the CAD group, the PCAT attenuation parameters of vessels with plaques were slightly higher than those of vessels without plaques (all P values >0.05). In receiver operating characteristic curve analysis, the FAIVMI model achieved an area under the curve (AUC) of 0.8123 for discriminating between patients with and without CAD, which was higher than those of the FAI120 kVp model (AUC =0.7444) and the λ model (AUC =0.7230). However, the combined model of FAIVMI, FAI120 kVp, and λ obtained the best performance (AUC =0.8296) of all the models. PCAT attenuation parameters obtained using dual-layer SDCT can aid in distinguishing patients with and without CAD. By detecting increases in PCAT attenuation parameters, it might be possible to predict the formation of atherosclerotic plaques before they appear.

Impact of glucose intolerance on the risk of carotid plaque instability assessed by MRI

Atherosclerotic disease is a complex multifactorial and systemic condition that affects arteries, including carotid arteries. Atherosclerotic plaque instability can predict plaque rupture, the major cause of ischemic strokes. Intraplaque hemorrhage (IPH) is an important contributor of plaque instability caused by the pro-inflammatory and hypoxic environment of the atherosclerotic plaque promoting the development of intraplaque neo-vessels. Metabolic disturbances are known to promote atherosclerotic plaque formation. Ischemic stroke prevalence is increase in patients with obesity and/or type-2 diabetes. The objective of the present study is to evaluate whether glucose intolerance can increase IPH in patients with carotid atherosclerosis. This study involves 57 patients with carotid atherosclerotic plaque with more than 50% stenosis, asymptomatic for more than 6 months, with no surgical indication for endarterectomy and no recent inflammatory disease. IPH was evaluated by magnetic resonance imaging (MRI). A score was defined for IPH (from 0: non hemorrhage to 3: large hemorrhage). Fasting blood glucose level was measured and patients were allocated in glucose tolerant (< 5,6 mM) or glucose intolerant (> 5,6 mM) groups. We showed that the score of IPH quantified by MRI was increased in the glucose intolerant subgroup of patients (0.39 vs. 0.84, P = 0.04). This result was confirmed by a higher prevalence of hemorrhagic plaque in the glucose intolerant group (73 vs. 41%, P < 0.05). Interestingly, the fasting blood glucose level is higher in patients with a hemorrhagic plaque than non-hemorrhagic plaque (5.6 vs. 6.9 mM, P = 0.04). Moreover, neither other known pro-atherogenic markers (LDL, cholesterol, TG) nor obesity seem to increase the degree of IPH. In conclusion, we demonstrated a strong impact of blood glucose level on the risk of plaque instability reflected by IPH. Further experiments are needed to strengthen the relation between type-2 diabetes, blood glucose level and IPH to understand the underlying mechanisms.

Homeobox A9 is a novel mediator of vascular smooth muscle cell phenotypic switching and proliferation by regulating methyl-CpG binding protein 2

Aberrant proliferation and phenotypic switching of vascular smooth muscle cells (VSMCs) are considered to be the main pathological processes of atherosclerotic plaque formation. Methyl-CpG binding protein 2 (MECP2) affects cell differentiation via modulating VSMC-specific gene expression and acts as a driver for the development of atherosclerosis (AS). Here, we aimed to elucidate the role of homeobox A9 (HOXA9) on aberrant VSMCs upon injury or AS, and whether HOXA9-mediated VSMC injury was associated with MECP2. Adeno-associated virus serotype 8-mediated knockdown of HOXA9 rescued aortic pathological injury of apolipoprotein E-deficient (ApoE−/−) mice fed a high-fat diet (HFD), characterized by the reduction of lipid accumulation and foam cell formation. Further in vitro evidence suggested that proliferation and migration of primary mouse VSMCs (mVSMCs) stimulated by oxidized low-density lipoprotein (ox-LDL) were inhibited after HOXA9 silencing. In addition, HOXA9 silencing blocked VSMC phenotypic switching from contractile to a pathological synthetic state. HOXA9 overexpression caused opposite alterations in ox-LDL-stimulated mVSMCs. Mechanistically, MECP2 was transcriptionally activated by HOXA9. Forced expression of MECP2 impaired the anti-proliferation, anti-migration, and phenotypic switching abilities of HOXA9 silencing in VSMCs upon ox-LDL stimulation. Collectively, our findings reveal that the role of HOXA9 in pathological vascular remodeling may attribute to transcriptional regulation of MECP2.

Sulforaphane Inhibits Foam Cell Formation and Atherosclerosis via Mechanisms Involving the Modulation of Macrophage Cholesterol Transport and the Related Phenotype

Sulforaphane (SFN), an isothiocyanate, is one of the major dietary phytochemicals found in cruciferous vegetables. Many studies suggest that SFN can protect against cancer and cardiometabolic diseases. Despite the proposed systemic and local vascular protective mechanisms, SFN’s potential to inhibit atherogenesis by targeting macrophages remains unknown. In this study, in high fat diet fed ApoE-deficient (ApoE−/−) mice, oral SFN treatment improved dyslipidemia and inhibited atherosclerotic plaque formation and the unstable phenotype, as demonstrated by reductions in the lesion areas in both the aortic sinus and whole aorta, percentages of necrotic cores, vascular macrophage infiltration and reactive oxygen species (ROS) generation. In THP-1-derived macrophages, preadministration SFN alleviated oxidized low-density lipoprotein (ox-LDL)-induced lipid accumulation, oxidative stress and mitochondrial injury. Moreover, a functional study revealed that peritoneal macrophages isolated from SFN-treated mice exhibited attenuated cholesterol influx and enhanced apolipoprotein A-I (apoA-I)- and high-density lipoprotein (HDL)-mediated cholesterol efflux. Mechanistic analysis revealed that SFN supplementation induced both intralesional and intraperitoneal macrophage phenotypic switching toward high expression of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1) and ATP-binding cassette subfamily A/G member 1 (ABCA1/G1) and low expression of peroxisome proliferator-activated receptor γ (PPARγ) and cluster of differentiation 36 (CD36), which was further validated by the aortic protein expression. These results suggest that the regulation of macrophages’ cholesterol transport and accumulation may be mainly responsible for SFN’s potential atheroprotective properties, and the regulatory mechanisms might involve upregulating ABCA1/G1 and downregulating CD36 via the modulation of PPARγ and Nrf2.

Five-Year Changes in Inflammatory, Metabolic, and Oxidative Biomarkers and 10-Year Cardiovascular Disease Incidence: The REGICOR Cohort Study.

Ischemic cardiovascular diseases (CVD) originate from an imbalance between atherosclerotic plaque formation, instability, and endothelial healing dynamics. Our aim was to examine the relationship between 5-year changes in inflammatory, metabolic, and oxidative biomarkers and 10-year CVD incidence in a population without previous CVD. This was a prospective cohort study of individuals aged 35-74 years (n = 419) randomly selected from 5263 REGICOR participants without CVD recruited in 2005. Biomarkers were measured at baseline and in 2010. Participants were followed up until 2020 for a composite CVD endpoint including coronary artery disease, stroke, and peripheral artery disease. We used Cox regression to analyze the effect of biomarker levels on the occurrence of the composite endpoint, adjusted for traditional CVD risk factors and baseline levels of each biomarker. Individuals with elevated IL-6 or insulin after 5 years had a higher independent risk of CVD at 10 years, compared to those with lower levels. Each rise of 1 pg/mL of IL-6 or 10 pg/mL of insulin increased the 10-year risk of a CVD event by 32% and 2%, respectively. Compared to a model with traditional CVD risk factors only, the inclusion of IL-6 and insulin improved continuous reclassification by 51%. Elevated serum levels of IL-6 and insulin were associated with a higher risk of CVD at 10 years, independently of traditional CVD risk factors.

SFRP4 Reduces Atherosclerosis Plaque Formation in ApoE Deficient Mice.

Secreted frizzled related protein 4 (SFRP4), a member of the SFRPs family, contributes to a significant function in metabolic and cardiovascular diseases. However, there is not enough evidence to prove the antiatherosclerosis effect of SFRP4 in ApoE knock-out (KO) mice. ApoE KO mice were fed a western diet and injected adenovirus (Ad)-SFRP4 through the tail vein for 12 weeks. Contrasted with the control cohort, the area of atherosclerotic plaque in ApoE KO mice overexpressing SFRP4 was reduced significantly. Plasma high-density lipoprotein cholesterol was elevated in the Ad-SFRP4 group. RNA sequence analysis indicated that there were 96 differentially expressed genes enriched in 10 signaling pathways in the mRNA profile of aortic atherosclerosis lesions. The analysis data also revealed the expression of a number of genes linked to metabolism, organism system, and human disease. In summary, our data demonstrates that SFRP4 could play an important role in improving atherosclerotic plaque formation in the aorta.

The cell origins of foam cell and lipid metabolism regulated by mechanical stress in atherosclerosis.

Atherosclerosis is an inflammatory disease initiated by endothelial activation, in which lipoprotein, cholesterol, extracellular matrix, and various types of immune and non-immune cells are accumulated and formed into plaques on the arterial wall suffering from disturbed flow, characterized by low and oscillating shear stress. Foam cells are a major cellular component in atherosclerotic plaques, which play an indispensable role in the occurrence, development and rupture of atherosclerotic plaques. It was previously believed that foam cells were derived from macrophages or smooth muscle cells, but recent studies have suggested that there are other sources of foam cells. Many studies have found that the distribution of atherosclerotic plaques is not random but distributed at the bend and bifurcation of the arterial tree. The development and rupture of atherosclerotic plaque are affected by mechanical stress. In this review, we reviewed the advances in foam cell formation in atherosclerosis and the regulation of atherosclerotic plaque and lipid metabolism by mechanical forces. These findings provide new clues for investigating the mechanisms of atherosclerotic plaque formation and progression.

San-wei-tan-xiang capsule attenuates atherosclerosis by increasing lysosomal activity in adipose tissue macrophages

Ethnopharmacological relevanceDyslipidemia is the leading risk factor of atherosclerosis (AS). Adipose tissue macrophages (ATMs) can regulate postprandial cholesterol levels via uptake and hydrolyzation of lipids and regulation of macrophage cholesterol efflux (MCE). San-wei-tan-xiang (SWTX) capsule, a Traditional Chinese medicine, exerts clinical benefits in patients with atherosclerotic cardiovascular diseases. Aim of the studyThis work is aimed to evaluate the chemical ingredients and mechanisms of SWTX in anti-AS. Materials and methodsThe chemical ingredients of SWTX identified by liquid chromatography coupled with tandem mass spectrometry were used for network pharmacological analysis. The atheroprotective function of SWTX was evaluated in ApoE−/− mice fed a cholesterol-enriched diet. ResultsThe chemical ingredients identified in SWTX were predicated to be important for lipid metabolism and AS. Animals studies suggested that SWTX effectively attenuated the atherosclerotic plaque growth, elevated postprandial HDL cholesterol levels, elevated the proportion of Tim4 and CD36-expressed ATMs, and upregulated the uptake of lipid and lysosomal activity in ATMs. SWTX-induced elevation of postprandial HDL cholesterol levels was dependent on increased lysosomal activity, since chloroquine, an inhibitor of lysosomal function, blocked the effect of SWTX. Lastly, some predicated bioactive compounds in SWTX can elevate lysosomal activity in vitro. ConclusionSWTX could attenuate atherosclerotic plaque formation by elevating lysosomal activity and enhancing MCE in ATMs.

Study on the effect of periodontitis on renal tissue in atherosclerotic mice

Periodontitis is immune inflammatory disease, atherosclerosis (AS) and chronic kidney disease (CKD) are two common systemic diseases. Periodontitis promotes AS and CKD, and CKD interacts with AS. The objective of this animal study was to evaluate the changes of kidney when periodontitis and atherosclerosis exist separately and the degenerative effects of periodontitis on the kidney in atherosclerotic mice. A total of 40 male Apoe-/- mice were randomly divided into four groups: control (NC), periodontitis (PD), AS and AS with PD (AS + PD). AS was induced by high-fat diet feeding, and PD was induced by injection of Porphyromonas gingivalis-Lipopolysaccharide (P.g-LPS) (endotoxin suspension) into the buccal side of mouse maxillary molars. The right maxilla of mice was scanned with micro-CT to evaluate alveolar bone loss; aortic tissue was stained with HE and Oil-Red O to evaluate arterial plaque formation; serum was collected to detect the changes of blood lipids and serum renal function parameters (blood urea nitrogen [BUN], serum creatinine [Scr]); renal histopathological changes were evaluated by HE staining (glomerular and tubular damage scores), PAS staining (glomerular Mesangial matrix index) and Masson staining (percentage of renal fibrosis area); qRT-PCR and ELISA were used to evaluate the expression of renal inflammatory cytokines (tumor necrosis factor-α, Interleukin-1β, neutrophil surface marker Ly6G). The amount of alveolar bone loss: PD group was significantly higher than NC group (p < .05); AS + PD group was higher than PD group, the difference was not statistically significant. Atherosclerotic plaque formation and serum lipid changes: AS group were significantly worse than NC group (p < .05), and AS + PD group were worse than AS group. The results of the corresponding qualitative and quantitative analyses of kidney tissue in experimental animals gradually deteriorated in the NC group, PD group, AS group and AS + PD group and worsened sequentially. Renal function parameters: the content of BUN in AS group was higher than that in PD group, the difference was not statistically significant; Scr in AS group was significantly higher than that in PD group (p < .05); the contents of BUN and Scr in AS + PD group were higher than those in AS group, the difference was not statistically significant. Glomerular and tubular damage scores: AS group were higher than PD group, the difference was not statistically significant; AS + PD group were significantly higher than AS group (p < .001). The ratio of glomerular mesangial matrix to glomerular area and the percentage of renal fibrosis area: AS group were significantly higher than PD group (p < .001), and AS + PD group were significantly higher than AS group (p < .001). Expression of inflammatory cytokines: AS group was higher than PD group, the difference was not statistically significant; AS + PD group was significantly higher than AS group (p < .05). Both PD and AS can aggravate the inflammatory stress of kidney tissue and cause the damage of kidney tissue, and the inflammatory increase and damage effect of AS is stronger; PD can promote kidney damage of atherosclerotic mice by aggravating the renal inflammation in atherosclerotic mice; renal function parameters were not completely synchronized with the changes of renal inflammation and histopathology in each group of mice; PD can promote AS, periodontal inflammation in mice with AS is more severe, and the special changes of blood lipids in mice with AS are closely related to the above results.

Dietary Evodiamine Inhibits Atherosclerosis-Associated Changes in Vascular Smooth Muscle Cells.

Evodia rutaecarpa (Juss.) Benth is a traditional Chinese medicine. The active ingredient, evodiamine, is a quinolone alkaloid and is found in Evodiae fructus. We investigated the effect of evodiamine on atherosclerosis using LDLR-/- mice fed on a high-fat diet and ox-LDL-induced MOVAS cell lines to construct mouse models and cell-line models. We report a significant reduction in atherosclerotic plaque formation in mice exposed to evodiamine. Our mechanistic studies have revealled that evodiamine can regulate the proliferation, migration, and inflammatory response of and oxidative stress in vascular smooth muscle cells by inhibiting the activation of the PI3K/Akt axis, thus inhibiting the occurrence and development of atherosclerosis. In conclusion, our findings reveal a role for evodiamine in the regulation of vascular smooth muscle cells in atherosclerosis, highlighting a potential future role for the compound as an anti-atherosclerotic agent.

Dry Fruits: Contain Chemicals to Keep Human Body Organs Healthy

Hyperglycemia, hypertension, and hyperlipidemia are known as metabolic syndrome. In presence of high levels of blood LDL particles, free radicals (ROS), macrophages, cytokines, platelets, histamine, 5HT get involved in LDL oxidation, leading to formation of atherosclerotic plaques which get deposited in endothelium of coronary arteries causing coronary artery disease (CAD). Psyllium fibers and Ficus Carica (FC) are being considered to be useful in treating hyperlipidemia, hypertension, and hyperglycemia. This study was conducted to determine hypotensive, hypolipidemic, and hypoglycemic potential of Psyllium husk and FC. Study was conducted in National hospital, Lahore, Pakistan from January to April 2020. Seventy five hyperlipidemic, hypertensive and diabetic patients were enrolled selected from the hospital with their consent. They were divided in three groups ie 25 patients in each group. Group-1 was on placebo, group-2 was on 15 grams psyllium husk to be used in three divided doses. Group-3 was on 12 grams FC to be taken in three divided doses daily for two months. Their Lipid profile, FBS, and hypertension were determined before and after two months therapy by these two agents. Two analyze data we used SPSS version 10.01.001 2015 and t test was applied to determine changes in the tested parameters. In two months therapy by psyllium husk in 24 patients TG, TC, LDL-c was reduced 21.48, 10.09, 24.17 mg/dl respectively. HDL was increased 12.38 mg/dl in this group. FBS reduction was 28.66 mg/dl, systolic BP 6.30, and diastolic BP reduced in this group 7.52 mm of mg. We then concluded that FC and PH have good enough potential to normalize lipid profile, reduce BP, and FBS in patients suffering from metabolic syndrome.

Can We Take Hyperuricemia as a Cardiovascular Risk?

Study conducted for Gouty arthritis at Pan Pakistan level in 2017 showed mean Uric Acid levels of 6.11±1.7mg/dl with frequency of hyperuricemia of 39% and having 27.9% male and 49.35 women respectively.1 This type of arthritis results from monosodium urate crystal deposition in and around the joints affecting big toe around the joints of hands and feet. Gout has been shown to increase with age and associated with diabetes &amp; hypertension.2,3 It has also been shown that hyperuricemia may be tied to increased risk of cardiovascular disease mortality.4 Hyperuricemia has also shown to increase atherosclerosis by systemic inflammation and oxidative stress. It also causes endothelial dysfunction and oxidation of lipoproteins with formation of atherosclerotic plaques.5 Recent studies have shown hypertension, coronary artery disease, cardiovascular diseases, vascular dementia and preclampsia to be associated with high uric acid levels. The studies include Rotterdam (Serum Uric Acid Level and Myocardial infraction),2 PUMA study (Serum Uric Acid levels with cardiovascular events and deaths in hypertensive patients),6 PAMELA study (increased risk of cardiovascular deaths with increase of every 1mg /dl).4 Three important co-relation i.e. Hyperuricemia, atherosclerosis and heart failure need bit of understanding. In one study elevated uric acid levels (75.5mg/dl) in 90% of adolescents with essential hypertension7 and lower uric acid levels in teens with either white coat or secondary hypertension.7 Secondly association between increased uric acid levels and coronary artery calcification has shown an independent risk factor for subclinical atherosclerosis in young adults.8 Similar association was also seen in atherosclerotic vulnerable carotid plaque.9 Thirdly in cardiovascular health study incidence of heart failure was 21% with Chronic hyperuricemia and 18% without.10 For management of hyperuricemia (gout) multidisciplinary approach involving primary care physicians, rheumatologist and cardiologists might improve associated cardiovascular diseases other than primary pathology. Drugs like non-steroidal anti-inflammatory drug (NSAIDS), steroids &amp; colchicine may improve Gout but improvement in cardiovascular comorbids are questionable.11,12 Though different epidemiological studies have shown hyperuricemia as an independent risk factor for cardiovascular diseases and hypertension. Further studies are needed that my elucidate association of increased uric acid level with cardiovascular diseases. Multidisciplinary approach, awareness and counselling patient will hopefully prevent onset of heart disease and improve survival outcomes for gouty patients.

Ficus deltoidea var. kunstleri Extract Administration in Hypercholesterolaemic, Atherosclerotic Rabbits: Effects on Organ Function, Morphology, and Atherosclerosis Development

Ficus deltoidea (FD) is used in traditional Malay medicine to treat various ailments and has been shown to be safe in toxicity studies. However, the information on the safety and efficacy of FD in the atherosclerosis-induced animal model is limited. This study aims to investigate the safety of FD var. kunstleri (FDK) extract on high cholesterol diet (HCD)-induced atherosclerotic rabbits and its efficacy in treating atherosclerosis. New Zealand White rabbits were randomly divided into two groups: G1 (1% HCD for 4 weeks) and G2 (1% HCD for 8 weeks). Each group was randomised into FDK700 (700 mg FDK/kg/day for G1 and G2), FDK800 (800 mg FDK/kg/day for G2), simvastatin (5 mg/kg/day) and placebo. The body weight, blood pressure, serum biochemistry and histopathological examination were obtained to assess any toxicity signs. Fasting lipid profile, soluble c-reactive protein (sCRP) level and atherosclerotic plaque formation were compared between treated and placebo groups to evaluate treatment efficacy. Results: No significant differences were observed in all safety parameters between the treated and placebo groups (p&lt;0.05). FDK treatment did not show significant differences in all parameters evaluated in both treatment arms. In conclusion, FDK extract up to 800 mg/kg is safe for use in atherosclerotic rabbits. It has neutral effects on lipid profile, inflammation and atherosclerosis formation.

SMTP-44D Inhibits Atherosclerotic Plaque Formation in Apolipoprotein-E Null Mice Partly by Suppressing the AGEs-RAGE Axis

SMTP-44D has been reported to have anti-oxidative and anti-inflammatory reactions, including reduced expression of receptor for advanced glycation end products (RAGE) in experimental diabetic neuropathy. Although activation of RAGE with its ligands, and advanced glycation end products (AGEs), play a crucial role in atherosclerotic cardiovascular disease, a leading cause of death in diabetic patients, it remains unclear whether SMTP-44D could inhibit experimental atherosclerosis by suppressing the AGEs-RAGE axis. In this study, we investigated the effects of SMTP-44D on atherosclerotic plaque formation and expression of AGEs in apolipoprotein-E null (Apoe-/-) mice. We further studied here whether and how SMTP-44D inhibited foam cell formation of macrophages isolated from Apoe-/- mice ex vivo. Although administration of SMTP-44D to Apoe-/- mice did not affect clinical or biochemical parameters, it significantly decreased the surface area of atherosclerotic lesions and reduced the atheromatous plaque size, macrophage infiltration, and AGEs accumulation in the aortic roots. SMTP-44D bound to immobilized RAGE and subsequently attenuated the interaction of AGEs with RAGE in vitro. Furthermore, foam cell formation evaluated by Dil-oxidized low-density lipoprotein (ox-LDL) uptake, and gene expression of RAGE, cyclin-dependent kinase 5 (Cdk5) and CD36 in macrophages isolated from SMTP-44D-treated Apoe-/- mice were significantly decreased compared with those from saline-treated mice. Gene expression levels of RAGE and Cdk5 were highly correlated with each other, the latter of which was also positively associated with that of CD36. The present study suggests that SMTP-44D may inhibit atherosclerotic plaque formation in Apoe-/- mice partly by blocking the AGEs-RAGE-induced ox-LDL uptake into macrophages via the suppression of Cdk5-CD36 pathway.

MiR-147a targets ZEB2 to regulate ox-LDL-induced monocyte adherence to HUVECs, atherosclerotic plaque formation, and stability in atherosclerosis

The mechanisms underlying atherosclerosis (AS) that seriously affect human health, such as those involved in endothelial cell injury and monocyte/macrophage aggregation and infiltration, have not been fully elucidated. To investigate these processes, we established human umbilical vein endothelial cells (HUVECs) injured by oxidized low-density lipoprotein (ox-LDL) to mimic AS invitro. Apolipoprotein E knockout (ApoE-/-) C57BL/6 mice were fed with a high-cholesterol diet to establish an AS model invivo. We detected HUVEC apoptosis, and apoptosis-related proteins by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide and lactate dehydrogenase, flow cytometry, and Western blot assays, respectively, and we observed monocytes (THP-1cells) adhering to HUVECs. Furthermore, miR-147a and its downstream target gene ZEB2 (zinc finger E-box binding homeobox 2) were predicted by bioinformatics analysis to be involved in AS, and their correlation was confirmed by several experiments. We determined the localization of miR-147a and ZEB2 within macrophages of AS mice by in situ hybridization and immunofluorescence. Atherosclerotic plaques in whole aortas were detected by histology observation. miR-147a attenuated adherence of monocytes to HUVECs and the upregulation of mononuclear chemotactic adhesion receptors in THP-1cells induced by ox-LDL-injured HUVEC supernatants through directly downregulating ZEB2 levels. Moreover, miR-147a influenced M1/M2 macrophage polarization from THP-1cells and the roles of their supernatants (THP-1cells) in HUVEC apoptosis. miR-147a targeted ZEB2 to impact lipid accumulation and atherosclerotic plaque formation through regulating M1/M2 polarization and macrophage adhesion in AS mice. In summary, miR-147a attenuates ox-LDL-induced adherence of monocytes to HUVECs and modulates atherosclerotic plaque formation and stability through targeting ZEB2 during AS.

The Role of Cytokines in Cholesterol Accumulation in Cells and Atherosclerosis Progression

Atherosclerosis is the most common cardiovascular disease and is the number one cause of death worldwide. Today, atherosclerosis is a multifactorial chronic inflammatory disease with an autoimmune component, accompanied by the accumulation of cholesterol in the vessel wall and the formation of atherosclerotic plaques, endothelial dysfunction, and chronic inflammation. In the process of accumulation of atherogenic lipids, cells of the immune system, such as monocytes, macrophages, dendritic cells, etc., play an important role, producing and/or activating the production of various cytokines-interferons, interleukins, chemokines. In this review, we have tried to summarize the most important cytokines involved in the processes of atherogenesis.

Comparing the effectiveness of pterostilbene and sitagliptin on modulating inflammatory levels and inducing autophagy to improve atherosclerosis outcome: A preclinical study in rabbits

Background: Inflammation is the key contributor to the development of atherosclerotic plague. This study aims to evaluate the protective and autophagy induction properties of pterostilbene and sitagliptin on modulating the degree of atherosclerosis in rabbit models treated with an atherogenic diet. Methods: 80 rabbits were randomly placed into one of four study groups (20 in each group): normal control diet (NC) fed normal diet for eight weeks, atherogenic control (AC) fed atherogenic diet for eight weeks, pterostilbene treated group (PT) fed atherogenic diet with pterostilbene (at 10 mg/kg/day) orally daily for eight weeks, and sitagliptin treated group (ST) fed atherogenic diet with sitagliptin (at 12 mg/kg/day) orally daily for eight weeks. Results: While serum lipids and F2-isoprostane were elevated significantly in the AC study cohort compared to NC study cohort, (P &lt; 0.001), both pterostilbene and sitagliptin supplementations provided significant improvements in serum lipid parameters and F2-isoprostane in the PT study cohort and ST study cohort, respectively, when compared to the AC study cohort, (P&lt;0.001). Total cholesterol, triglycerides and LDL levels were significantly reduced among the PT and ST study cohorts as compared to the AC study cohort. This was coupled with a significant rise in LC3B levels (marker of tissue autophagy) among the PT study cohort and the ST study cohort, as compared to the AC study cohort, (P &lt; 0.001). The RNA expression of mTORC1 was reduced significantly at both PT study cohort and ST study cohort, (P&lt;0.001). Pterostilbene supplementation induced a significant reduction in tissue expression of PI3K and AKT, (P&lt;0.01), while sitagliptin induced significant reduction in 5’ adenosine monophosphate-activated protein kinase (AMPK) levels, (P&lt;0.001). Conclusions: The results indicate that pterostilbene and/or sitagliptin supplementation can significantly improve the outcome of atherosclerosis due to their effects on the inflammatory pathways which hinder the progression of atherosclerotic plaque formation.