Atherosclerosis Plaque Research Articles

Exercise-mediated adaptations in vascular function and structure: Beneficial effects in coronary artery disease.

Exercise exerts direct effects on the vasculature via the impact of hemodynamic forces on the endothelium, thereby leading to functional and structural adaptations that lower cardiovascular risk. The patterns of blood flow and endothelial shear stress during exercise lead to atheroprotective hemodynamic stimuli on the endothelium and contribute to adaptations in vascular function and structure. The structural adaptations observed in arterial lumen dimensions after prolonged exercise supplant the need for acute functional vasodilatation in case of an increase in endothelial shear stress due to repeated exercise bouts. In contrast, wall thickness is affected by rather systemic factors, such as transmural pressure modulated during exercise by generalized changes in blood pressure. Several mechanisms have been proposed to explain the exercise-induced benefits in patients with coronary artery disease (CAD). They include decreased progression of coronary plaques in CAD, recruitment of collaterals, enhanced blood rheological properties, improvement of vascular smooth muscle cell and endothelial function, and coronary blood flow. This review describes how exercise via alterations in hemodynamic factors influences vascular function and structure which contributes to cardiovascular risk reduction, and highlights which mechanisms are involved in the positive effects of exercise on CAD.

Prevalence of Microalbuminuria and Its Association with Subclinical Carotid Atherosclerosis in Middle Aged, Nondiabetic, Low to Moderate Cardiovascular Risk Individuals with or without Hypertension.

Microalbuminuria is closely associated with the risk of cardiovascular disease and all-cause mortality in the general population. Less is known about its relationship with subclinical atherosclerosis. We aimed to assess the prevalence of microalbuminuria and its relationship with subclinical atherosclerosis in middle-aged, nondiabetic, apparently healthy individuals (N = 187; 40.1% men, 59.9% women; aged 35–55 years) as well as to evaluate its potential associations with established risk modifiers, especially with the presence of carotid plaque. Clinical and laboratory parameters, the estimated 10-year fatal cardiovascular risk (SCORE), as well as circulating, functional (flow mediated vasodilation, ankle-brachial index, augmentation index, and pulse wave velocity), and morphological markers (mean carotid intima–media thickness, and carotid plaque) of subclinical atherosclerosis were analysed in group with vs. without microalbuminuria. Microalbuminuria was present in 3.8% of individuals with SCORE risk 0.43 ± 0.79%. Functional markers predominated in both groups. Carotid intima–media thickness (mean ± SD) in both groups was in range: 0.5–0.55 ± 0.09–0.14 mm. Carotid plaque was more frequent in group with (14.3%) vs. without (4.4%) microalbuminuria. Microalbuminuria had no statistically significant effect on most markers of subclinical atherosclerosis, but the increasing value of microalbuminuria was significantly associated with the occurrence of carotid plaque (p = 0.035; OR = 1.035; 95% CI = 1.002–1.07). Additional multiple logistic regression analysis, where variables belonged to microalbuminuria, number of risk factors, and family history, finally showed only two variables: microalbuminuria (p = 0.034; OR = 1.04; 95%CI = 1.003–1.09) and the number of risk factors (p = 0.006; OR = 2.15; 95% CI = 1.24–3.73) with independent and significant impact on the occurrence of carotid plaque. Our results may indicate an association of microalbuminuria with the presence of carotid atherosclerotic plaque; in addition, microalbuminuria and the number of risk factors appear to be possible predictors of the carotid plaque occurrence. Monitoring microalbuminuria may improve the personalized cardiovascular risk assessment in nondiabetic, low-to-moderate cardiovascular risk individuals with or without hypertension.

HAP-Multitag, a PET and Positive MRI Contrast Nanotracer for the Longitudinal Characterization of Vascular Calcifications in Atherosclerosis.

Vascular microcalcifications are associated with atherosclerosis plaque instability and, therefore, to increased mortality. Because of this key role, several imaging probes have been developed for their in vivo identification. Among them, [18F]FNa is the gold standard, showing a large uptake in the whole skeleton by positron emission tomography. Here, we push the field toward the combined anatomical and functional early characterization of atherosclerosis. For this, we have developed hydroxyapatite (HAP)-multitag, a bisphosphonate-functionalized 68Ga core-doped magnetic nanoparticle showing high affinity toward most common calcium salts present in microcalcifications, particularly HAP. We characterized this interaction in vitro and in vivo, showing a massive uptake in the atherosclerotic lesion identified by positron emission tomography (PET) and positive contrast magnetic resonance imaging (MRI). In addition, this accumulation was found to be dependent on the calcification progression, with a maximum uptake in the microcalcification stage. These results confirmed the ability of HAP-multitag to identify vascular calcifications by PET/(T1)MRI during the vulnerable stages of the plaque progression.

The Anti-Atherosclerotic Action of FFAR4 Agonist TUG-891 in ApoE-Knockout Mice Is Associated with Increased Macrophage Polarization towards M2 Phenotype.

Background: Over the past few years, a better understanding of the biology of G-protein coupled receptors (GPRs) has led to the identification of several receptors as novel targets for free fatty acids (FFAs). FFAR4 has received special attention in the context of chronic inflammatory diseases, including atherosclerosis, obesity and NAFLD, through to its anti-inflammatory effect. Methods: The present study investigates the influence of prolonged treatment with TUG-891-FFAR4 agonist on the development of atherosclerosis plaque in apoE-knockout mice, using morphometric and molecular methods. Results: TUG-891 administration has led to the reduction of atherosclerotic plaque size and necrotic cores in an apoE-knockout mice model. TUG-891-treated mice were administered subcutaneously at a dose of 20 mg/kg three times a week for 4 months. The FFAR4 agonist reduced the content of pro-inflammatory M1-like macrophages content in atherosclerotic plaques, as evidenced by immunohistochemical phenotyping and molecular methods. In atherosclerotic plaque, the population of smooth muscle cells increased as evidenced by α-SMA staining. We observed changes in G-CSF and eotaxin markers in the plasma of mice; changes in the levels of these markers in the blood may be related to macrophage differentiation. Importantly, we observed a significant increase in M2-like macrophage cells in atherosclerotic plaque and peritoneum. Conclusions: Prolonged administration of TUG-891 resulted in significant amelioration of atherogenesis, providing evidence that the strategy based on macrophage phenotype switching toward an M2-like activation state via stimulation of FFAR4 receptor holds promise for a new approach in the prevention or treatment of atherosclerosis.

Multi-Feature Fusion Method for Identifying Carotid Artery Vulnerable Plaque

PurposeVulnerable plaque of carotid atherosclerosis is prone to rupture, which can easily lead to acute cardiovascular and cerebrovascular accidents. Accurate identification of the vulnerable plaque is a challenging task, especially on limited datasets. MethodsThis paper proposes a multi-feature fusion method to identify high-risk plaque, in which three types of features are combined, i.e. global features of carotid ultrasound images, echo features of regions of interests (ROI) and expert knowledge from ultrasound reports. Due to the fusion of three types of features, more critical features for identifying high-risk plaque are included in the feature set. Therefore, better performance can be achieved even on limited datasets. ResultsFrom testing all combinations of three types of features, the results showed that the accuracy of using all three types of features is the highest. The experiments also showed that the performance of the proposed method is better than other plaque classification methods and classical Convolutional Neural Networks (CNNs) on the Plaque dataset. ConclusionThe proposed method helped to build a more complete feature set so that the machine learning models could identify vulnerable plaque more accurately even on datasets with poor quality and small scale.

Current Status of Primary, Secondary, and Tertiary Prevention of Coronary Artery Disease.

Fifty percent of all death from cardiovascular diseases is due to coronary artery disease (CAD). This is avoidable if early identification is made. Preventive health care has a major role in the fight against CAD. Atherosclerosis and atherosclerotic plaque rupture are involved in the development of CAD. Modifiable risk factors for CAD are dyslipidemia, diabetes, hypertension, cigarette smoking, obesity, chronic renal disease, chronic infection, high C-reactive protein, and hyperhomocysteinemia. CAD can be prevented by modification of risk factors. This paper defines the primary, secondary, and tertiary prevention of CAD. It discusses the mechanism of risk factor-induced atherosclerosis. This paper describes the CAD risk score and its use in the selection of individuals for primary prevention of CAD. Guidelines for primary, secondary, and tertiary prevention of CAD have been described. Modification of risk factors and use of guidelines for prevention of CAD would prevent, regress, and slow down the progression of CAD, improve the quality of life of patient, and reduce the health care cost.

Status of biomarkers for the identification of stable or vulnerable plaques in atherosclerosis.

Atherosclerosis is a systemic inflammation of the arteries characterized by atherosclerotic plaque due to the accumulation of lipids, inflammatory cells, apoptotic cells, calcium and extracellular matrix (ECM) proteins. Stable plaques present a chronic inflammatory infiltration, whereas vulnerable plaques present an 'active' inflammation involved in the thinning of the fibrous cap that predisposes to plaque rupture. Several complex biological cellular processes lead plaques to evolve from stable to vulnerable predisposing them to rupture and thrombosis. In this review, we analyze some emerging circulating biomarkers related to inflammation, ECM and lipid infiltration, angiogenesis, metalloproteinases and microRNA (miRNA), as possible diagnostic and prognostic indicators of plaque vulnerability.

Specific Targeting MRI of Chitosan Oligosaccharide Modified Fe3O4 Nanoprobe on Macrophage and the Inhibition of Macrophage Foaming Induced by ox-LDL

Atherosclerosis (AS) is a primary cause of morbidity and mortality all over the world. Molecular imaging techniques can enable early localization and diagnosis of atherosclerosis plaques. Recent newly developed chitooligosaccharides (CSO) is considered to be capable of target mannose receptors on the surface of macrophages and to inhibit foam cell formation. Here we present a targeting magnetic resonance imaging (MRI) nanoprobe, which was successfully constructed with polyacrylic acid (PAA) modified nanometer iron oxide (Fe3O4) as the core, and coating with CSO molecules, possessing the abilities of targeted MRI and specifically inhibition of the formation of foamy macrophages in the atherosclerotic process. The experimental results showed that the distributions of PAA-Fe3O4 and CSO-PAA-Fe3O4 were uniform and the corresponding sizes were about 5.93 nm and 8.15 nm, respectively. The Fourier transform infrared spectra (FTIR) testified the CSO was coupled with PAA-Fe3O4 successfully. After coupled with CSO, the r1 of PAA-Fe3O4 was increased from 5.317 mM s-1 to 6.147 mM s-1, indicating their potential as MRI contrast agent. Oil Red O staining and total cholesterols (TC) determination showed that CSO-PAA-Fe3O4 could significantly inhibit the foaming process of RAW264.7 cells induced by oxidatively modified low density lipoprotein (ox-LDL). In vitro cellular MRI displayed that, compared with PAA-Fe3O4,CSO-PAA-Fe3O4 could lower the T1 relaxation time of RAW264.7 cells better. In summary, construction of CSO-PAA-Fe3O4 nanoprobe in this study could realize the targeted MRI of macrophages and inhibition of ox-LDL induced macrophage foaming process. This will provide a new avenue in the diagnosis and treatment of AS.

MMP-10 rs17435959 Polymorphism is Associated with the Formation and Stability of Carotid Atherosclerosis Plaque: A Case-Control Study

BackgroundMatrix metalloproteinase 10 (MMP-10) has a close relationship with carotid atherosclerosis (CAS) and cerebral infarction. The MMP-10 rs17435959 polymorphism causes a leucine to valine transition at codon 4 in exon 1 of the MMP-10 gene and may have functional effects. ObjectivesTo investigate the relationship between the MMP-10 rs17435959 polymorphism and the formation and stability of CAS plaques. Materials and MethodsThe present case-control study contains 738 visitors who came to our health examination center for the first time. According to the carotid ultrasound examinations, visitors were classified into the vulnerable plaque group (41–86 years old, 141 male, 105 female), the stable plaque group (41–86 years old, 141 male, 105 female) and the no plaque group (41–85 years old, 141 male, 105 female). All visitors in the three groups were sex- and- age-matched, and cardiovascular and cerebrovascular diseases were absent. The polymorphism was genotyped by real-time polymerase chain reaction- restriction. ResultsCompared to the GG genotype, the frequency of the CC and CG genotypes was significantly more common in the vulnerable plaque group than in the no plaque group (18.7% vs. 7.7%, unadjusted P = 0.002). Moreover, compared to the G allele, the frequency of the C allele was significantly more common in the vulnerable plaque group than in the no plaque group (10.4% vs. 3.9%, unadjusted P = 0.000) and in the vulnerable plaque group than in the stable plaque group (10.4% vs. 5.1%, unadjusted P = 0.008). Binary logistic regression showed that the CC and CG genotype was independent risk factor for the formation (P = 0.019, OR = 1.961, 95% CI [1.117, 3.444]) and vulnerability (P = 0.035, OR = 1.842, 95% CI [1.045, 3.247]) of CAS plaques. Moreover, individuals who have the C allele showed a higher level of fibrinogen, which was an independent risk factor for the formation of CAS plaques (P = 0.000, OR = 2.425, 95% CI [1.475, 3.985]). ConclusionsThe rs17435959 polymorphism was associated with the formation and vulnerability of CAS plaques. Individuals who had variant-type MMP-10 showed higher levels of fibrinogen, which promoted the formation of CAS plaques.

IL-37 polymorphisms are associated with the presence of coronary artery disease and cardiovascular risk factors. The GEA mexican study.

Background and Aims: Interleukin 37 (IL-37) is an anti-inflammatory cytokine involved in the regulation of cholesterol homeostasis. IL-37 expression has been reported in foam cells located in the atherosclerosis plaques. The aim of the present study was to evaluate the association of the IL-37 polymorphisms with premature coronary artery disease (pCAD), and with cardiovascular risk factors.

Single-cell analysis uncovers osteoblast factor GDF10 as mediator of vascular smooth muscle cell phenotypic modulation associated with plaque rupture in human carotid artery disease

Background and Aims: Vascular smooth muscle cells (VSMC) are not terminally differentiated and undergo complex phenotypic changes during atherosclerosis. However, the complex heterogeneity of VSMC and how VSMC de-differentiation affects human carotid artery disease (CAD) risk has not been clearly established. The aim of the present study is to comprehensively characterized the transcriptomic profile of phenotypically modulated VSMC and to identify mediators of VSMC transition to osteogenic like cells with likely detrimental role for atherosclerosis plaque stability.

A fuzzy-logic-based fault detection system for medical Internet of Nano Things

In this paper, a fuzzy-logic-based fault detection system is designed for a medical Internet of Nano Things architecture. The goal of this system is to detect the root cause and severity of the faults occurred in the in-body nanonetwork. Since nanomachines have very limited capabilities, the sampled data from the in-body nanonetwork is sent to cloud servers by means of an on-body micro-gateway. The fuzzy fault detection system was designed based on two well-known methods including Mamdani and Takagi–Sugeno–Kang (TSK) fuzzy systems. The performance of the proposed approach is evaluated on a theoretical model of medical in-body nanonetwork from the literature through in silico study. This nanonetwork includes eleven types of nanomachines which cooperate with each other within the arterial wall and interact with low-density lipoprotein (LDL), drug and signaling molecules in order to prevent the formation and development of Atherosclerosis plaques. Any fault in these nanomachines can highly take negative effect on treatment efficiency. The results of computer simulation and comparative study on 37 atherosclerosis patients demonstrate how the proposed approach could successfully detect the root cause and severity of the faults occurred in the nanonetwork.

Exercise training mitigates ER stress and UCP2 deficiency-associated coronary vascular dysfunction in atherosclerosis

Endoplasmic reticulum (ER) stress and uncoupling protein-2 (UCP2) activation are opposing modulators of endothelial dysfunction in atherosclerosis. Exercise reduces atherosclerosis plaques and enhances endothelial function. Our aim was to understand how exercise affects ER stress and UCP2 activation, and how that relates to endothelial dysfunction in an atherosclerotic murine model. Wild type (C57BL/6, WT) and apolipoprotein-E-knockout (ApoEtm1Unc, ApoE KO) mice underwent treadmill exercise training (EX) or remained sedentary for 12 weeks. Acetylcholine (ACh)-induced endothelium-dependent vasodilation was determined in the presence of an eNOS inhibitor (L-NAME), UCP2 inhibitor (genipin), and ER stress inducer (tunicamycin). UCP2, ER stress markers and NLRP3 inflammasome signaling were quantified by western blotting. p67phox and superoxide were visualized using immunofluorescence and DHE staining. Nitric oxide (NO) was measured by nitrate/nitrite assay. ACh-induced vasodilation was attenuated in coronary arterioles of ApoE KO mice but improved in ApoE KO-EX mice. Treatment of coronary arterioles with L-NAME, tunicamycin, and genipin significantly attenuated ACh-induced vasodilation in all mice except for ApoE KO mice. Exercise reduced expression of ER stress proteins, TXNIP/NLRP3 inflammasome signaling cascades, and Bax expression in the heart of ApoE KO-EX mice. Further, exercise diminished superoxide production and NADPH oxidase p67phox expression in coronary arterioles while simultaneously increasing UCP2 expression and nitric oxide (NO) production in the heart of ApoE KO-EX mice. Routine exercise alleviates endothelial dysfunction in atherosclerotic coronary arterioles in an eNOS, UCP2, and ER stress signaling specific manner, and resulting in reduced TXNIP/NLRP3 inflammasome activity and oxidative stress.

Balectin-3 affects the stability of vulnerable coronary atherosclerosis plaques by targeted modulation of lncARSR

To investigate the mechanism by which galectin-3 (Gal-3) affects the stability of vulnerable coronary atherosclerosis plaques through long non-coding RNA ARSR (lncARSR). Male BALB/c mice were randomly divided into normal diet group, high-fat diet group, high-fat diet+lncARSR inhibitor group (n=20). The high-fat diet contained 15% fat and 0.25% cholesterol, and lncARSR inhibitor was injected intravenously at 50 nmol/L every other day. After 12 weeks of high-fat diet feeding and treatment, the mice were euthanized for analyzing coronary atherosclerosis and plaque damage using Sudan IV and oil red O staining. The protein expressions of Gal-3 and ARSR in the coronary artery of the mice were analyzed with Western blotting, and the expressions of PI3K and Akt were detected with immunohistochemistry. The coronary artery tissues were harvested from normal mice for cell culture, and the isolated cells were transfected with a Gal-3 mimic or a Gal-3 inhibitor. At 24 h after the transfection, dual luciferase reporter gene assay was performed to determine the target relationship between Gal-3 and lncARSR; the mRNA expressions of tumor necrosis factor (TNF-α), interleukin-β (IL-β) and IL-6 in the transfected cells were detected with RT-qPCR. The positively stained areas by Sudan IV and red oil O and the protein expression of lncARSR were the lowest in normal diet group and the highest in high-fat diet group (P < 0.05). The protein expression of PI3K and Akt and the mRNA expression of TNF-α, IL-β and IL-6 in high-fat diet group were higher than those in normal diet group. The protein expression of PI3K and Akt and the mRNA expression of TNF-α, IL-β and IL-6 in high-fat diet+ lncARSR inhibitor group were significantly lower than those in high-fat diet group (P < 0.05). In the cell experiment, the activity of WT-lncARSR was significantly higher in Gal-3 mimic transfection group than in the control group and Gal-3 inhibition group (P=0.026), and was the lower in Gal-3 inhibition group than in the control group (P=0.017). Gal-3 and lncARSR are overexpressed in coronary atherosclerosis. Through a mechanism for targeted inhibition of lncARSR, Gal-3 regulates the PI3K/ Akt signaling pathway to suppress inflammation and thus regulate the stability of vulnerable coronary atherosclerosis plaques.

A lipid droplets specific probe for imaging of atherosclerosis and fibrocalcific bicuspid aortic valves

Lipid droplets (LDs) have been regarded as potential targets for the diagnosis of cardiovascular diseases. However, conventional LDs imaging strategies suffer from invasive sample collection and tedious histological preparations. Although fluorescence imaging has showed good performance in LDs imaging, ideal probes are still in urgent demand. In this work, fluorescent probe, namely, MeTTI was prepared for LDs visualization in cells and tissues. MeTTI exhibited low fluorescence quantum yield in dimethyl sulfoxide (Φf = 0.2 %), while the value in the solid state was obvious increased (Φf = 3.5 %), suggesting the aggregation-induced emission (AIE) property of MeTTI. Meanwhile, MeTTI would form excimers (emission ranged 750–850 nm) in water, and it showed good biocompatibility. Moreover, MeTTI exhibited impressive fluorescence emission enhancement in oleic acid (Φf = 34.7 %) compared to that in water (Φf = 1.11 %), which can specifically stain LDs in cells, atherosclerosis (AS) plaques and human fibrocalcific bicuspid aortic valves (FBAV) with high ratio of signal to noise. In addition, MeTTI could also monitor the dynamic changes of LDs in cells. These results were expected to provide a simple tool for studying the pathogenesis of cardiovascular diseases with lipid deposition and triggering enthusiasm to develop new imaging techniques for the diagnosis of these diseases.

Anti-inflammatory cytokine IL10 loaded cRGD liposomes for the targeted treatment of atherosclerosis

Aim Atherosclerosis (AS) is one of the main causes of cardiovascular disease which might lead to myocardial infarction or stroke and further leads to fatality. Method In this study, we have designed an anti-inflammatory cytokine interleukin-10 (IL10) delivery system to effectively alleviate the inflammation of atherosclerosis plaque. The targeted delivery of IL10 to the atherosclerotic plaques was achieved by cRGD conjugated liposomes (IL10-cRGD-Lip). Results The IL10-cRGD-Lip of size 179.4 ± 10.91 nm having PDI 0.14 ± 0.04 with a surface charge of +18.34 ± 1.36 mV was prepared. The in-vitro analysis clearly suggests that IL10-cRGD-Lip sustains the release of IL10 and could significantly reduce ROS and NO. The immuno-staining results revealed that IL-1β and TNF-α were down-regulated after the treatment with IL10-cRGD-Lip in Lipopolysaccharide (LPS) stimulated RAW 264.7 cells. Conclusion the in-vitro results clearly suggest that anti-inflammatory cytokine IL10 could be used for the cure of inflammatory maladies including atherosclerosis.

18F-ASEM Imaging for Evaluating Atherosclerotic Plaques Linked to α7-Nicotinic Acetylcholine Receptor.

BackgroundAtherosclerosis is a chronic vascular inflammatory procedure alongside with lipid efflux disorder and foam cell formation. α7-Nicotinic acetylcholine receptor (α7nAChR) is a gated-calcium transmembrane channel widely expressed in neuron and non-neuron cells, such as monocytes and macrophages, activated T cells, dendritic cells, and mast cells. 18F-ASEM is an inhibitor targeted to α7nAChR that had been successfully applied in nervous system diseases. Previous studies had highlighted that α7nAChR was related to the emergency of vulnerable atherosclerotic plaques with excess inflammation cells. Thus, 18F-ASEM could be a complementary diagnostic approach to atherosclerotic plaques.Materials and MethodsThe synthesis of ASEM precursor and 18F-labeling had been performed successfully. We had established the ApoE–/– mice atherosclerotic plaques model (fed with western diet) and New Zealand rabbits atherosclerotic models (balloon-sprained experiment and western diet). After damage of endothelial cells and primary plaque formation, 18F-ASEM imaging of atherosclerotic plaques linked to α7nAChR had been conducted. In vivo micro-PET/CT imaging of ApoE–/– mice and the control group was performed 1 h after injection of 18F-ASEM (100–150 μCi); PET/CT imaging for rabbits with atherosclerotic plaques and control ones was also performed. Meanwhile, we also conducted CT scan on the abdominal aorta of these rabbits. After that, the animals were sacrificed, and the carotid and abdominal aorta were separately taken out for circular sections. The paraffin-embedded specimens were sectioned with 5 μm thickness and stained with hematoxylin–eosin (H&E) and oil red.ResultsIn vivo vessel binding of 18F-ASEM and α7nAChR expression in the model group with atherosclerosis plaques was significantly higher than that in the control group. PET/CT imaging successfully identified the atherosclerotic plaques in ApoE–/– mice and model rabbits, whereas no obvious signals were detected in normal mice or rabbits. Compared with 18F-FDG, 18F-ASEM had more significant effect on the early monitoring of inflammation in carotid atherosclerotic plaques of ApoE–/– mice and model rabbits. 18F-ASEM had relatively more palpable effect on the imaging of abdominal aorta with atherosclerosis in rabbits. H&E and oil red staining identified the formation of atherosclerotic plaques in model animals, which provided pathological basis for the evaluation of imaging effects.ConclusionWe first confirmed 18F-ASEM as radiotracer with good imaging properties for precise identification of atherosclerotic diseases.

Effects of Hamo NK hard capsule on experimental atherosclerosis model

This study evaluated the effects of Hamo NK hard capsule on athresclerosis using experimental atherosclerosis model. NewZealand White rabbits were fed a high-fat diet (HFD) containing cholesterol and peanut oil. The animals received oral administration of HFD and Hamo NK hard capsule at two doses of 0.126 and 0.378 g/kg bw/day for 8 consecutive weeks. Blood samples were collected for analyis of biochemical parameters at before treatment, week 4 and week 8. Histopathology assessments of the aortic artery and liver were carried out at the end of the experiment. Hamo NK was effective in reducing serum triglyceride level after 8 weeks of the experiment. In addition, Hamo NK at two doses of 0.126 g/kg b.w and 0.378 g/kg b.w for 8 consecutive weeks did not affect the cholesterol, LDL-C and HDL-C concentrations induced by a HFD. Hamo NK at the dose of 0.126 g/kg bw/day was not only able to decrease significant aortic surface lesions but also capable of managing atherosclerosis plaques formation in aorta; whereas theses activities were not notiaceable at the dose of 0.378 g/kg b.w.

Traditional Chinese Medication Tongxinluo Attenuates Lipidosis in Ox-LDL-Stimulated Macrophages by Enhancing Beclin-1-Induced Autophagy.

Tongxinluo (TXL), a traditional Chinese medication, plays a key role in the formation and progression of plaques in atherosclerosis. The formation of foam cells by macrophages accelerates the destabilisation of plaques. In previous research, we had found that TXL significantly inhibits ox-LDL-induced apoptosis in macrophages in vitro by improving the dissociation of the Beclin-1-Bcl-2 complex. Therefore, here, we explored the effect of TXL on lipid metabolism in macrophages and the mechanism involved. To evaluate the role of TXL in atherosclerotic plaques, we construct the atherosclerotic animal model with lentiviral injection and performed immunofluorescence staining analysis in vivo. Western blot, immunofluorescence staining and microscopy were performed to elucidate the mechanism underlying TXL-mediated regulation of autophagy in THP-1 macrophages in vitro. Immunofluorescence assay revealed that TXL treatment inhibited lipid deposition in advanced atherosclerotic plaques. In vitro TXL treatment inhibited lipid deposition in THP-1 macrophages by enhancing autophagy via Beclin-1. TXL reversed the high expression of class I histone deacetylases (HDACs) induced by ox-LDL (p < 0.05). Compared with the TXL + ox-LDL group, TXL failed to promote intracellular lipid droplet decomposition after the addition of the histone deacetylase agonist. We found that TXL attenuates the accumulation of lipids in macrophage by enhancing Beclin-1-induced autophagy, and additionally, it inhibits the inhibitory effect of class I HDAC on the expression of Beclin-1.

A radioiodinated FR-β-targeted tracer with improved pharmacokinetics through modification with an albumin binder for imaging of macrophages in AS and NAFL.

The formation of advanced plaques, which is characterized by the uninterrupted aggregation of macrophages with high expression of folate receptor-β (FR-β), is observed in several concomitant metabolic syndromes. The objective of this study was to develop a novel FR-β-targeted single-photon emission computed tomography (SPECT) radiotracer and validate its application to the noninvasive detection of atherosclerosis (AS) plaque and non-alcoholic fatty liver (NAFL). Two radioiodinated probes, [131I]IPBF and [131I]IBF, were developed, and cell uptake studies were used to identify their specific targets for activated macrophages. Biodistribution in normal mice was performed to obtain the pharmacokinetic information of the probes. Apolipoprotein E knockout (ApoE-/-) mice with atherosclerotic aortas were induced by a high-fat and high-cholesterol (HFHC) diet. To investigate the affinity of radiotracers to FR-β, Kd values were determined using in vitro assays. In addition, the assessments of the aorta in the ApoE-/- mice at different stages were performed using in vivo SPECT/CT imaging, and the findings were compared by histology. Both [131I]IPBF and [131I]IBF were synthesized with > 95% radiochemical purity and up to 3MBq/nmol molar activity. In vitro assay of [131I]IPBF showed a moderate binding affinity to plasma proteins and specific uptake in activated macrophages. The prolonged blood elimination half-life (t1/2z) of [131I]IPBF (8.14h) was observed in a pharmacokinetic study of normal mice, which was significantly longer than that of [131I]IBF (t1/2z = 2.95h). As expected, the Kd values of [131I]IPBF and [131I]IBF in the Raw 264.7 cells were 43.94 ± 9.83nM and 61.69 ± 15.19nM, respectively. SPECT imaging with [131I]IPBF showed a high uptake in advanced plaques and NAFL. Radioactivity in excised aortas examined by ex vivo autoradiography further confirmed the specific uptake of [131I]IPBF in high-risk AS plaques. In summary, we reported a proof-of-concept study of an albumin-binding folate derivative for macrophage imaging. The FR-β-targeted probe, [131I]IPBF, significantly prolongs the plasma elimination half-life and has the potential for the monitoring of AS plaques and concomitant fatty liver.