The associations between TMAO-related metabolites and blood lipids and the potential impact of rosuvastatin therapy.

BACKGROUND: Trimethylamine N-oxide (TMAO)-related metabolites are associated with the pathogenesis of atherosclerotic cardiovascular disease (ASCVD) and are known to disrupt lipid metabolism. The aims of this study were to evaluate the associations between TMAO-related metabolites and blood lipids and determine how lowering the lipid profile via rosuvastatin therapy influences TMAO-related metabolites. METHODS: A total of 112 patients with suspected ASCVD were enrolled in this study. The levels of plasma TMAO-related metabolites, including TMAO, choline, carnitine, betaine, and γ-butyrobetaine (GBB), were analyzed by stable isotope dilution liquid chromatography-tandem mass spectrometry (LC/MS/MS) before and after rosuvastatin therapy in all patients. Statistical methods were used to detect the associations between TMAO-related metabolites and blood lipids and determine how rosuvastatin therapy alters the levels of these metabolites. RESULTS: A significant positive correlation was found between TMAO and triglycerides (TG) (r = 0.303, P < 0.05). Furthermore, significant negative correlations were found between TMAO and high-density lipoprotein cholesterol (HDL-c) and between betaine and low-density lipoprotein cholesterol (LDL-c) (r = - 0.405 and - 0.308, respectively, both P < 0.01). Compared to baseline, significantly lower TMAO levels and higher carnitine, betaine and GBB levels were observed after rosuvastatin therapy, while the lipids decreased significantly (P < 0.05). The significant correlation between TMAO and TG or between betaine and LDL-c disappeared after rosuvastatin therapy (r = 0.050 and - 0.172, respectively, both P > 0.05). However, a significantly positive association between carnitine and TC and a negative association between carnitine and LDL-c or between betaine and TG were found after adjustment for sex, age, body mass index (BMI) and lipids (P < 0.05). CONCLUSIONS: This study suggests that TMAO-related metabolites are significantly associated with blood lipids, although some of them are changed postrosuvastatin therapy. Lower TMAO and higher TMAO precursors were observed after rosuvastatin therapy compared to baseline. This study indicates that elevated TMAO precursors after rosuvastatin therapy and their potential impact on ASCVD should be considered in the clinic.

Soluble receptor for advanced glycation end-products enhanced the production of IFN-γ through the NF-κB pathway in macrophages recruited by ischemia/reperfusion.

The current study investigated the role of sRAGE in the production of IFN­Î³ in macrophages with I/R treatment. The number of macrophages in myocardial tissues treated with I/R with or without sRAGE was determined via immunohistochemical staining. Proliferative activity of macrophages was analyzed by a 5­BrdU incorporation assay. Differentiation of macrophages was detected via immunofluorescence staining of iNOS (M1 macrophage marker). IFN­Î³ production, due to sRAGE stimulation, in Raw 264.7 macrophages and the NF­κB signaling pathway were measured using western blotting. A ChIP assay was used to examine the interactions between NF­κB and the promoter of IFN­Î³. The results showed that the number of macrophages in I/R­treated myocardial tissues was increased following sRAGE infusion. Proliferation of macrophages was increased significantly in the presence of sRAGE; after I/R treatment, the cells preferred to differentiate into M1 macrophages. IFN­Î³ expression in Raw 264.7 macrophages was suppressed by an NF­κB inhibitor (Bay117082) but enhanced by sRAGE, with or without I/R treatment. Furthermore, sRAGE increased the phosphorylation of IκB, IKK and NF­κB, as well as the translocation of NF­κB into the nucleus of Raw 264.7 macrophages, with or without I/R treatment. ChIP results showed that sRAGE promoted NF­κB binding to the promoter of IFN­Î³ in Raw 264.7 macrophages. Therefore, the findings of the present study indicated that sRAGE protected the heart from I/R injuries, which might be mediated by promoting infiltration and the differentiation of macrophages into M1, which would then synthesize and secrete IFN­Î³ through activating the NF­κB signaling pathway.

Interferon-γ mediates the protective effects of soluble receptor for advanced glycation end-product in myocardial ischemia/reperfusion.

The ubiquitin-proteasome system (UPS) is essential for protein degradation and plays critical roles in myocardial ischemia/reperfusion (MI/R) injuries. Previous studies have demonstrated that the soluble receptor for advanced glycation end-product (sRAGE) inhibited MI/R-induced apoptosis by upregulating proteasome subunits. However, the mechanism remains unknown. An MI/R model was established by left anterior descending (LAD) coronary artery ligation in mice. Recombinant sRAGE protein or saline was injected intramyocardially with or without neutralizing interferon-γ (IFN-γ) antibody injected intraperitoneally before ligation. In cardiomyocytes, ischemia was simulated with "ischemia buffer" and sRAGE was overexpressed by adenovirus. Adenovirus expressing the interference RNA of ß5i was used to knockdown ß5i in cardiomyocytes. IFN-γ was induced by sRAGE both in sham and MI/R mice. Blockade of IFN-γ using IFN-γ antibody abolished the rescue effects of sRAGE for cardiac dysfunction, infarct size and apoptosis provoked by MI/R. Blockade of IFN-γ reversed the upregulation of ß1i and ß5i expression induced by sRAGE during MI/R in heart, accompanied by decreasing chymotrypsin-like proteasome activity. In addition, IFN-γ antibody abolished the suppressing effect of sRAGE on MI/R-induced p38 and c-Jun N-terminal kinase (JNK) activation, as well as p53 expression, both in vivo and in vitro. However, knockdown of ß5i abolished the antiapoptosis effect of sRAGE during hypoxia/reoxygenation (H/R) in vitro, accompanied by decreased degradation of p53. Our data suggest a novel mechanism for sRAGE in preventing MI/R-induced apoptosis in heart: sRAGE inhibits MI/R-induced apoptosis in cardiomyocytes by degrading p53 by ß5i subunit that is increased via upregulation of IFN-γ.

Soluble receptor for advance glycation end-products inhibits ischemia/reperfusion-induced myocardial autophagy via the STAT3 pathway.

The pathogenesis of myocardial ischemia/reperfusion (I/R) is poorly understood, but recent evidence suggests that autophagy plays crucial roles in I/R injuries. Soluble receptor for advanced glycation end-products (sRAGE) exerts protective effects during I/R by decreasing cardiac apoptosis, which is mediated via increasing the ubiquitin proteasome system (UPS) and signal transducer and activator of transcription 3 (STAT3). The present study examined the effects and mechanisms of sRAGE on I/R-triggered cardiac autophagy. I/R was performed in mice or primary neonatal cardiomyocytes with or without sRAGE administration or overexpression. Cardiac function and infarct size were detected in mouse hearts. Apoptosis, autophagy and autophagy-related signaling pathways were detected in mouse hearts and cardiomyocytes. The results demonstrated that sRAGE significantly improved cardiac function and reduced infarct size during I/R in mice. sRAGE inhibited I/R-induced apoptosis, which correlated with a reduction in autophagy-associated proteins, including ATG7, Beclin-1 and microtubule-associated protein 1 light chain 3 (LC3). sRAGE reduced autophagosome formation during I/R in vivo and in vitro. sRAGE significantly activated STAT3, but not mammalian target of rapamycin (mTOR), during I/R in vivo and in vitro, and suppression of STAT3 abolished the sRAGE inhibition of autophagy during I/R in vitro. Activation of autophagy using ATG7 overexpression with an adenovirus significantly abolished the sRAGE-induced reduction of cardiac apoptosis during I/R. These results suggest that sRAGE inhibits I/R injuries in the heart via a decrease in autophagy, a process that is dependent on STAT3 activation.

Endogenous cholesterol ester hydroperoxides modulate cholesterol levels and inhibit cholesterol uptake in hepatocytes and macrophages.

Dysregulation of cholesterol metabolism represents one of the major risk factors for atherosclerotic cardiovascular disease (CVD). Oxidized cholesterol esters (oxCE) in low-density lipoprotein (LDL) have been implicated in CVD but the underlying mechanisms remain poorly defined. We use a targeted lipidomic approach to demonstrate that levels of oxCEs in human plasma are associated with different types of CVD and significantly elevated in patients with myocardial infarction. We synthesized a major endogenous cholesterol ester hydroperoxide (CEOOH), cholesteryl-13(cis, trans)-hydroperoxy-octadecadienoate (ch-13(c,t)-HpODE) and show that this endogenous compound significantly increases plasma cholesterol level in mice while decrease cholesterol levels in mouse liver and peritoneal macrophages, which is primarily due to the inhibition of cholesterol uptake in macrophages and liver. Further studies indicate that inhibition of cholesterol uptake by ch-13(c,t)-HpODE in macrophages is dependent on LXRα-IDOL-LDLR pathway, whereas inhibition on cholesterol levels in hepatocytes is dependent on LXRα and LDLR. Consistently, these effects on cholesterol levels by ch-13(c,t)-HpODE are diminished in LDLR or LXRα knockout mice. Together, our study provides evidence that elevated plasma cholesterol levels by CEOOHs are primarily due to the inhibition of cholesterol uptake in the liver and macrophages, which may play an important role in the pathogenesis of CVD.

Change in lipoprotein-associated phospholipase A2 and its association with cardiovascular outcomes in patients with acute coronary syndrome.

Lipoprotein-associated phospholipase A2 (Lp-PLA2) probably plays an important role in the development of acute coronary syndrome (ACS). However, alterations of Lp-PLA2 levels during ACS and its association with cardiovascular outcome are unclear. Our aim was to investigate the change in Lp-PLA2 and its association with cardiovascular outcome in patients with ACS.A total of 79 patients with ACS came from the coronary care unit (CCU) between June 1, 2015 and August 31, 2016 in this longitudinal study. Serum levels of Lp-PLA2, troponin I, and creatine kinase isoenzymes MB (CK-MB) were measured at admission, on the first morning (D1), on the second morning of hospitalization (D2), and on the last second morning before discharge (D4). The patients were followed up till November 30, 2016. The primary outcomes were cardiovascular death and cardiovascular rehospitalization. Kaplan-Meier analysis and Cox proportional hazard models were used to identify risk factors for poor outcome in patients with ACS.All patients were followed up for 10.6 ±â€Š4.7 months. The patients were divided into 2 groups according to the median of Lp-PLA2: lower Lp-PLA2 group and higher Lp-PLA2 group. Elevated levels of Lp-PLA2 significantly decreased during the early phases of ACS in higher Lp-PLA2 group. And Lp-PLA2 level increased at first and then decreased in lower Lp-PLA2 group. Kaplan-Meier analysis showed that patients with elevated Lp-PLA2 had a lower cardiovascular event-free survival (log-rank χ = 4.736, P = .030) than those with lower Lp-PLA2. Cox regression analysis indicated that high Lp-PLA2 level (hazard ratio [HR] = 1.005, 95% confidence interval [CI] = 1.002-1.008, P = .003), time delay from symptom onset to admission (HR = 1.088, 95% CI = 1.038-1.139, P < .001) independently predicted cardiovascular event in patients with ACS after adjusting for potential confounders.Serum level of Lp-PLA2 altered considerably during the early phase of ACS and increased Lp-PLA2 independently predicted cardiovascular outcome in patients with ACS after adjustment for potential confounders.

Assessment of patients' warfarin knowledge and anticoagulation control at a joint physician- and pharmacist-managed clinic in China.

PURPOSE: Warfarin is a widely used anticoagulant with a narrow therapeutic index, and it requires close monitoring and adequate patient education. We aimed to assess the knowledge level regarding warfarin therapy among its users and to identify the factors that significantly influence anticoagulation control. PATIENTS AND METHODS: Patients attending the Warfarin Clinic at the Beijing Tiantan Hospital were enrolled in this study. Patients' knowledge on warfarin was assessed using a validated Anticoagulation Knowledge Assessment (AKA) questionnaire. Patients' responses to each question were analyzed to identify areas of improvement in current warfarin education. International normalized ratio (INR) control was defined by the time in therapeutic range (TTR) calculated using the Rosendaal method. Spearman correlation analysis was used to investigate the association between TTR and the independent variables. RESULTS: A total of 65 patients were enrolled in this study. Eleven questions were answered correctly by <50% of the patients. A total of 858 INR results were recorded; 432 INR values (50.3%) reached the predefined goals, and the mean TTR was 49.8%±24.8%. There were significant associations between TTR and patients' AKA scores (R=0.356, P=0.004) and between TTR and patients' educational levels (R=0.339, P=0.006). No significant association was observed between other factors (age and duration of anticoagulation) and TTR. The INR outcome measure was positively associated with patients' knowledge on warfarin and their educational levels. CONCLUSION: Areas for improvement in patient education have been identified, and processes for educational modification are currently in development.

Liraglutide downregulates hepatic LDL receptor and PCSK9 expression in HepG2 cells and db/db mice through a HNF-1a dependent mechanism.

BACKGROUND: Proprotein convertase subtilisin/kexin type 9 (PCSK9), a major regulator of cholesterol homeostasis, is associated with glucose metabolism. Liraglutide, a glucagon-like peptide-1 receptor agonist, can increase insulin secretion in a glucose-dependent manner and lower blood glucose. We aimed to investigate the relationship between liraglutide and PCSK9. METHODS: At the cellular level, the expressions of PCSK9 and hepatocyte nuclear factor 1 alpha (HNF1α) protein in HepG2 cells stimulated by liraglutide was examined using Western blot. Seven-week old db/db mice and wild type (WT) mice were administered either liraglutide (200 µg/kg) or equivoluminal saline subcutaneously, twice daily for 7 weeks. Fasting glucose level, food intake and body weight were measured every week. After the 7-week treatment, the blood was collected for lipid and PCSK9 levels detection and the liver was removed from the mice for oil red O staining, immunohistochemical analysis, immunofluorescence test and Western bolt. RESULTS: Firstly, liraglutide suppressed both PCSK9 and HNF1α expression in HepG2 cells in a time and concentration dependent manner. Secondly, liraglutide induced weight loss in WT and db/db mice, decreased serum PCSK9, glucose and lipid levels and improved hepatic accumulation in db/db but not WT mice. Thirdly, liraglutide reduced both hepatic PCSK9 and low-density lipoprotein receptor (LDLR) expression with a decrease in HNF1α in db/db mice but not in WT mice. CONCLUSIONS: Liraglutide suppressed PCSK9 expression through HNF1α-dependent mechanism in HepG2 cells and db/db mice, and decreased LDLR possibly via PCSK9-independent pathways in db/db mice.

Young Paradoxical Stroke Treated Successfully with Mechanical Thrombectomy Using Solitaire and Transcatheter Closure of Patent Foramen Oval.

Paradoxical embolization is the mechanism for patent foramen ovale (PFO)-associated cryptogenic stroke and transcatheter closure of PFO may prevent recurrent ischemic stroke. Mechanical thrombectomy is promising to treat acute ischemic stroke due to high rates of reperfusion and reduced intracranial hemorrhage complications. We report the case of a 27-year-old woman with a massive cerebral infarction but no evidence for any atherosclerosis, who received an urgent mechanical thrombectomy with a Solitaire device. In order to ascertain the etiology of stroke, transcranial Doppler (TCD) and transesophageal echocardiograph (TEE) were conducted. TCD showed severe right-to-left shunting (shower effect) after Valsalva maneuver and bubble test and TEE identified a PFO. Therefore, the patient had suffered a paradoxical stroke associated with PFO. After two weeks of the stroke onset, transcatheter PFO closure with Cardio-O-Fix occluder was also performed successfully. During 1-year of follow-up, no recurrence of stroke occurred. Our case demonstrates that mechanical thrombectomy using a Solitaire device and transcatheter PFO closure can be safely and successfully performed to treat acute paradoxical stroke and prevent its recurrence.

Electrocardiogram in patients with acute inferior myocardial infarction due to occlusion of circumflex artery.

To investigate the diagnostic value of electrocardiographic (ECG) ST-segment in acute inferior myocardial infarction (AIMI) caused by the left circumflex branch (LCX).A total of 240 clinical cases with AIMI in our hospital were retrospectively analyzed. All of them had received percutaneous coronary intervention (PCI) within 12 hours after symptom onset. The clinical features, ECG manifestations, and coronary artery lesion characteristics of the patients were collected.The right coronary artery (RCA) was shown to be the infarct-related artery (IRA) in 177 patients, while LCX was responsible for AIMI in 63 cases. There was no significant difference in the risk factors of coronary heart disease (CHD) (P > .05 for all) between the 2 groups. ST-segment elevation in lead II, III, and AVF could be found in all patients. Moreover, ST-segment depression in lead I (STD I), ST-segment elevation in lead III (STE III), STE III-STE II, STE AVF, STD AVL, STD AVL-STD I and STE v6 lead ST-segment deviation exhibited significant difference in 2 groups (P < .05 for all). The changes of STD I, STE III < STEII, STD AVL < STD I could discriminate between LCX and RCA in AIMI patients with high sensitivity and specificity.ECG may be an effective tool to predict the IRA in patient with AIMI.

15-oxoeicosatetraenoic acid mediates monocyte adhesion to endothelial cell.

BACKGROUND: A great number of studies reported that 12/15-lipoxygenase (12/15-LO) played an important role in atherosclerosis. And its arachidonic acid(AA) metabolite, 15(S)-hydroperoxy-5,8,11,13-(Z,Z,Z,E)-eicosatetraenoic acid (15(S)-HETE), is demonstrated to mediate endothelial dysfunction. 15-oxo-5,8,11,13-(Z,Z,Z,E)-eicosatetraenoic acid (15-oxo-ETE) was formed from 15-hydroxyprostaglandin dehydrogenase (PGDH)-mediated oxidation of 15(S)-HETE. However, relatively little is known about the biological effects of 15-oxo-ETE in cardiovascular disease. Here, we explore the likely role of 15-lipoxygenase (LO)-1-mediated AA metabolism,15-oxo-ETE, in the early pathogenesis of atherosclerosis. METHODS: The 15-oxo-ETE level in serum was detected by means of liquid chromatography and online tandem mass spectrometry (LC-MS/MS). And the underlying mechanisms were illuminated by molecular techniques, including immunoblotting, MTT assay, immunocytochemistry and Immunohistochemistry. RESULTS: Increased 15-oxo-ETE level is found in in patients with acute myocardial infarction (AMI). After 15-oxo-ETE treatment, Human umbilical vein endothelial cells (HUVECs) showed more attractive to monocytes, whereas monocyte adhesion is suppressed when treated with PKC inhibitor. In ex vivo study, exposure of arteries from C57 mice and ApoE-/-mice to 15-oxo-ETE led to significantly increased E-selectin expression and monocyte adhesion. CONCLUSIONS: This is the first report that 15-oxo-ETE promotes early pathological process of atherosclerosis by accelerating E-selectin expression and monocyte adhesion. 15-oxo-ETE -induced monocyte adhesion is partly attributable to activation of PKC.

Association Between Helicobacter Pylori Infection and Long-term Outcome in Patients with Drug-eluting Stent Implantation.

To investigate the relationship between Helicobacter pylori (Hp) infection and the long-term outcome in acute coronary syndrome (ACS) patients with drug-eluting stent (DES) implantation and so as to explore the significance of Hp eradication therapy in preventing major adverse cardiac events (MACE) and upper gastrointestinal bleeding (UGIB). 539 ACS patients with DES implantation from January 1, 2010 to December 31, 2012 were analyzed. All the patients were divided into two groups according to the result of 13C urea breath test. 253 patients with Hp infection were put into group A (Hp+), and 286 cases without Hp infection were put into group B (Hp-). Demographic data was collected and all patients went through biochemical indicators and other routine blood examinations. We explored the correlations of Hp infection with MACE and UGIB after 3 to 5 years of follow-up using survival analysis. Survival analysis showed that Hp infection was a predictor of MACE and UGI. Sub-group analysis showed that patients with Hp eradication therapy had no relationship with MACE but had a lower rate of UGIB than those without Hp eradication therapy.

Comprehensive metabolomics identified lipid peroxidation as a prominent feature in human plasma of patients with coronary heart diseases.

Coronary heart disease (CHD) is a complex human disease associated with inflammation and oxidative stress. The underlying mechanisms and diagnostic biomarkers for the different types of CHD remain poorly defined. Metabolomics has been increasingly recognized as an enabling technique with the potential to identify key metabolomic features in an attempt to understand the pathophysiology and differentiate different stages of CHD. We performed comprehensive metabolomic analysis in human plasma from 28 human subjects with stable angina (SA), myocardial infarction (MI), and healthy control (HC). Subsequent analysis demonstrated a uniquely altered metabolic profile in these CHD: a total of 18, 37 and 36 differential metabolites were identified to distinguish SA from HC, MI from SA, and MI from HC groups respectively. Among these metabolites, glycerophospholipid (GPL) metabolism emerged as the most significantly disturbed pathway. Next, we used a targeted metabolomic approach to systematically analyze GPL, oxidized phospholipid (oxPL), and downstream metabolites derived from polyunsaturated fatty acids (PUFAs), such as arachidonic acid and linoleic acid. Surprisingly, lipids associated with lipid peroxidation (LPO) pathways including oxidized PL and isoprostanes, isomers of prostaglandins, were significantly elevated in plasma of MI patients comparing to HC and SA, consistent with the notion that oxidative stress-induced LPO is a prominent feature in CHD. Our studies using the state-of-the-art metabolomics help to understand the underlying biological mechanisms involved in the pathogenesis of CHD; LPO metabolites may serve as potential biomarkers to differentiation MI from SA and HC.

Vascular response profiles following a nano polymer-free sirolimus-eluting stent implantation assessed by optical coherence tomography in a porcine model.

Optical coherence tomography (OCT) is a high resolution imaging modality and has been considered as the ideal tool for the evaluation of neointimal tissue and vascular responses following stent implantation. However, vascular response profiles following the implantation of a novel nano polymer-free sirolimus-eluting stent (SES) assessed by OCT has not been fully investigated. Therefore, the aim of the present study was to determine the effects of a nano polymer-free SES on neointimal formation using OCT. A total of 16 nano polymer-free SESs were implanted in the coronary arteries of 8 pigs. At 3 (n=4) or 6 months (n=4), the animals were euthanized following OCT evaluation and the stented arterial segments were analyzed by histological analysis. Neointimal area, thickness and burden were evaluated by OCT. In addition, strut-associated inflammation, stent endothelialization and arterial injury were investigated by histomorphological analysis. OCT examination showed that at 6 months, neointimal thickness (193.3±109.5 vs. 167.2±119.7 µm, P=0.023) and neointimal burden (29.3±14.3 vs. 24.8±17.4%, P=0.006) significantly increased compared with at 3 months. Histomorphological analysis indicated that the endothelialization score was significantly greater at 6 months compared with at 3 months (2.85±0.36 vs. 2.52±0.60, P<0.001). However, at 3 months, nano polymer-free SES showed a significantly higher inflammatory score [0 (0, 1) vs. 0 (0, 0), P<0.001] compared with at 6 months. In conclusion, nano polymer-free SES achieves endothelialization at 3 months; however, neointimal proliferation is more significant at 6 months and may be attributed to strut-associated inflammation.

Trimethylamine N-oxide in atherogenesis: impairing endothelial self-repair capacity and enhancing monocyte adhesion.

Several studies have reported a strong association between high plasma level of trimethylamine N-oxide (TMAO) and atherosclerosis development. However, the exact mechanism underlying this correlation is unknown. In the present study, we try to explore the impact of TMAO on endothelial dysfunction. After TMAO treatment, human umbilical vein endothelial cells (HUVECs) showed significant impairment in cellular proliferation and HUVECs-extracellular matrix (ECM) adhesion compared with control. Likewise, TMAO markedly suppressed HUVECs migration in transwell migration assay and wound healing assay. In addition, we found TMAO up-regulated vascular cell adhesion molecule-1 (VCAM-1) expression, promoted monocyte adherence, activated protein kinase C (PKC) and p-NF-κB. Interestingly, TMAO-stimulated VCAM-1 expression and monocyte adherence were diminished by PKC inhibitor. These results demonstrate that TMAO promotes early pathological process of atherosclerosis by accelerating endothelial dysfunction, including decreasing endothelial self-repair and increasing monocyte adhesion. Furthermore, TMAO-induced monocyte adhesion is partly attributable to activation of PKC/NF-κB/VCAM-1.

Identification of a novel series of anti-inflammatory and anti-oxidative phospholipid oxidation products containing the cyclopentenone moiety in vitro and in vivo: Implication in atherosclerosis.

Oxidative stress and inflammation are two major contributing factors to atherosclerosis, a leading cause of cardiovascular disease. Oxidation of phospholipids on the surface of low density lipoprotein (LDL) particles generated under oxidative stress has been associated with the progression of atherosclerosis, but the underlying molecular mechanisms remain poorly defined. We identified a novel series of oxidation products containing the cyclopentenone moiety, termed deoxy-A2/J2-isoprostanes-phosphocholine, from 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine in vivo using mass spectrometry and by comparison to a chemically synthesized standard. Transcriptomic analysis (RNA-seq) demonstrated that these compounds affected >200 genes in bone marrow-derived macrophages, and genes associated with inflammatory and anti-oxidative responses are among the top 5 differentially expressed. To further investigate the biological relevance of these novel oxidized phospholipids in atherosclerosis, we chemically synthesized a representative compound 1-palmitoyl-2-15-deoxy-δ-12,14-prostaglandin J2-sn-glycero-3-phosphocholine (15d-PGJ2-PC) and found that it induced anti-inflammatory and anti-oxidant responses in macrophages through modulation of NF-κB, peroxisome proliferator-activated receptor γ (PPARγ), and Nrf2 pathways; this compound also showed potent anti-inflammatory properties in a mice model of LPS-induced systematic inflammatory response syndrome. Additionally, 15d-PGJ2-PC inhibited macrophage foam cell formation, suggesting a beneficial role against atherosclerosis. These properties were consistent with decreased levels of these compounds in the plasma of patients with coronary heart disease compared with control subjects. Our findings uncovered a novel molecular mechanism for the negative regulation of inflammation and positive enhancement of anti-oxidative responses in macrophages by these oxidized phospholipids in LDL in the context of atherosclerosis.

Coronary Plaque Characterization Assessed by Optical Coherence Tomography and Plasma Trimethylamine-N-oxide Levels in Patients With Coronary Artery Disease.

Optical coherence tomography (OCT) has been considered as the ideal tool for the evaluation of atherosclerotic plaques. Circulating trimethylamine-N-oxide (TMAO), which is a metabolite of the dietary lipid phosphatidylcholine, has recently been linked to elevated coronary artery disease (CAD) risk. The objective of the study was to investigate the relation between circulating TMAO level and coronary plaque vulnerability assessed by OCT in patients with CAD. A total of 26 patients with CAD were recruited to assess coronary plaque using OCT and measure plasma TMAO level. According to plaque rupture status, patients were divided into plaque rupture group (n = 12) and nonplaque rupture group (n = 14). Plasma TMAO level was significantly higher in patients with plaque rupture than in those with nonplaque rupture (8.6 ± 4.8 µmol/L vs 4.2 ± 2.4 µmol/L, p = 0.011). Moreover, positive correlations between plasma TMAO level and lipid arc (r = 0.43, p = 0.031), lipid volume index (r = 0.39, p = 0.048) were also observed. In conclusion, circulating TMAO level may reflect coronary plaque vulnerability and progression.

Soluble receptor for advanced glycation end-products protects against ischemia/reperfusion-induced myocardial apoptosis via regulating the ubiquitin proteasome system.

AIM: Apoptosis participated in the pathological process of myocardial ischemia/reperfusion (I/R) injury. Previous studies have reported that endogenous substance sRAGE protect against I/R injury through inhibiting myocardial apoptosis. But the mechanisms are currently unknown. Prior work has demonstrated that ubiquitin proteasome system (UPS) dysfunction is closely related to apoptosis. We explored the potential role of UPS in the effect of sRAGE inhibition on I/R-induced myocardial apoptosis. METHODS AND RESULTS: Adult male C57BL mice treated with sRAGE (100µg/day, i.p.) or saline were performed to ligate left anterior descending coronary artery (LAD) as an in vivo model. As an in vitro model, primary murine cardiomyocytes pretreated with sRAGE or sRAGE-containing adenovirus were simulated I/R by "ischemia buffer". The TUNEL and caspase-3 activity were assessed. Also the activity and expression of proteasome were detected. sRAGE decreased the number of TUNEL-positive cardiomyocytes and caspase-3 activity, however, the inhibition of sRAGE on I/R-induced apoptosis was abolished by proteasome inhibitor Bortezimb (BTZ). sRAGE inhibited the decreased proteasome activity, also the reduction in protein and gene levels of ß1i and ß5i following I/R. Suppression of STAT3 blocked the inhibition of sRAGE on apoptosis induced by I/R. The chromatin immunoprecipitation (CHIP) results confirmed that sRAGE promoted activating STAT3 binding to ß1i and ß5i promoter. CONCLUSIONS: Our data suggest that the inhibition of sRAGE on I/R-induced apoptosis is associated with activation and expression of proteasome, including improved proteasome activity and elevated ß1i and ß5i expression mediated by STAT3 activation. We predict that sRAGE is a novel intervention to target UPS activation for preventing and treating myocardial apoptosis.

Relationship of coronary collateral circulation with eosinophils in patients with unstable angina pectoris.

BACKGROUND: Eosinophils (EOS) have been associated with prognosis of patients with coronary artery disease, and those who showed plenitudinous coronary collateral circulation (CCC) often have good clinical consequences. However, the relationship between EOS and CCC was seldom reported. OBJECTIVE: To investigate the relationship between EOS and CCC development in patients with unstable angina pectoris (UAP). METHODS: The study population consisted of 502 consecutive patients with UAP who underwent coronary angiography and coronary stenosis ≥80%. CCC was graded according to the Rentrop grading system of 0-3. Rentrop grades of 0 and 1 indicated low-grade CCC group, whereas grades 2 and 3 indicated high-grade CCC group. RESULTS: The EOS was significantly higher in the high-grade CCC group compared with the low-grade CCC group. In multiple logistic regression analysis, EOS (odds ratio: 1.969; 95% confidence interval [CI]: 1.210-3.3205; P=0.006) and neutrophil count (odds ratio: 0.757; 95% CI: 0.584-0.981; P=0.035) were predictors of high-grade CCC development. EOS of >0.12×10(9)/L could independently predict high-grade CCC with 72.5% sensitivity and 58.4% specificity (area under the curve: 0.681; 95% CI: 0.632-0.729). CONCLUSION: EOS were associated with high-grade CCC in patients with UAP with coronary stenosis ≥80%. Increased EOS count may play an important role in the development of CCC in patients with UAP.

[Carotid plaque characteristics detected with 3.0T high resolution nuclear magnetic resonance imaging in patients with coronary artery disease].

OBJECTIVE: To assess the value of detecting the compositional features of carotid atherosclerotic plaques by 3.0T high resolution magnetic resonance imaging (MRI) in patients with coronary artery disease (CAD). METHODS: Consecutive 104 patients with coronary atherosclerosis confirmed by coronary angiography were prospectively recruited from January 2013 to January 2015 in Tiantan hospital. All patients were imaged with 3.0T high resolution MRI system. After exclusion patients with poor image quality, 97 patients were divided into 3 groups according to the degree of coronary artery stenosis: coronary atherosclerosis group (coronary stenosis between 1%-49%, n=16); single-vessel lesion group (single vessel lesion with stenosis between 50%-100%, n=48); multi-vessel lesion group (two or three vessel lesions with stenosis between 50%-100% or left main stem disease, n=33). The prevalence of total carotid plaque, calcified plaque, lipid-rich necrotic core, intra-plaque hemorrhage, plaque ulcer and rupture were compared among 3 groups. RESULTS: The prevalence of total carotid plaque (81.3%(13/16), 72.9%(35/48), and 93.9%(31/33)) and calcified plaque (50.0%(8/16), 35.4%(17/48), and 42.4%(14/33)) were similar among the 3 groups (both P>0.05). The prevalence of carotid lipid-rich necrotic core in coronary atherosclerosis group was significantly lower than in single-vessel lesion group (18.8%(3/16) vs. 64.6%(31/48), P<0.01) and multi-vessel lesion group(18.8%(3/16) vs. 69.7%(23/33), P<0.01), but there was no significant difference between single-vessel lesion group and multi-vessel lesion group(P>0.05). Intra-plaque hemorrhage was detected in 2 patients of multi-vessel lesion group. There was no plaque ulcer or rupture in this cohort. CONCLUSION: Carotid plaque features are associated with the severity of coronary atherosclerosis in CAD patients.