Aortic Endothelial Dysfunction Research Articles

Olive cake reduces blood pressure, oxidative stress, aortic endothelial dysfunction and vascular remodeling, in dexamethasone-induced hypertensive rats

BACKGROUND/AIM: Hypertension is a global public health problem and a leading cause of cardiovascular morbidity and mortality. In this paper, we study the effect of olive cake (OC) on blood pressure, endothelial dysfunction, redox status and vascular remodeling in dexamethasone-induced hypertensive (HT) rats. METHODS: HT rats were divided into two groups fed standard diet supplemented (HT-OC) or not (HT) with OC at 7.5% for 28 days. A control (C) was submitted to standard diet for the same experimental period. RESULTS: Systolic, diastolic and mean blood pressures were higher in the HT vs C and decreased in HT-OC vs HT. Aortic nitric oxide value was decreased in the HT vs the C and increased in HT-OC vs HT. Aortic lipid and protein oxidation products were higher in the HT than C and lower in the HT-OC vs HT. Aortic antioxidant enzymes activities were reduced in HT than control and increased in the HT-OC vs HT. The aortic wall thickness, medial cross-sectional area, media to lumen ratio and the number of VSMCs were higher in the HT and the OC has regressed vascular redemptions. CONCLUSION: In hypertensive rats, OC may alleviate blood pressure and arterial remodeling by suppressing oxidative stress, increasing antioxidant activity, improving endothelial function, preventing smooth muscle proliferation and thickening of the tunica media.

Attenuation of streptozotocin induced high fat diet exacerbated dyslipidemia mediated hepatic and aortic injuries in male pigs by camel milk

ABSTRACT Dyslipidemia, diabetes, and the upregulation of receptors for oxidized low-density lipoprotein all contribute to hepatic and aortic endothelial dysfunction. The current research examined how camel milk could possibly decrease induced Streptozotocin-high fat diet-aggravated dyslipidemia-mediated damage in male pig hepatic and aortic tissues. Twenty-five pigs were separated into five groups of five pigs each, with five non-atherosclerotic pigs in the control group and twenty atherosclerotic pigs in the experimental groups. Groups 1 and 2 received distilled water as the standard control and atherosclerotic control groups, respectively, while Groups 3 and 4 received camel milk at 250 mL/day and 500 mL/day, respectively, and Group 5 received Metformin at 500 mg/day. The experiment lasted ten weeks. After 10 weeks, all of the pigs were euthanized. The lipid profile and liver enzyme levels were determined from the analysis of blood serum and the histology of the liver and aortic. When pigs administered, camel milk were compared to atherosclerotic control pigs, there was a significant decrease (p 0.05) in cholesterol, triglyceride, low density lipoprotein (LDL), and very low-density lipoprotein (VLDL) levels, while a significant increase in high density lipoprotein levels was observed in pigs that received camel milk. Also, pigs in groups that received camel milk had their liver enzyme parameters (ALP, AST, ALT, GGT, LDH, and CK-MB) significantly lowered when compared with atherosclerotic control pigs. Similarly, treatment with camel milk ameliorates aortic tissues from degradation and reduces histological liver steatosis, and it also decreases the expression of LOX-1 mRNA in the aorta of atherosclerotic pigs.

Oleocanthal supplemented diet improves renal damage and endothelial dysfunction in pristane-induced systemic lupus erythematosus in mice

Systemic lupus erythematosus (SLE) is a multiorgan disorder with a deregulated immune-inflammatory response. Nutritional therapy has been considered a promising approach to SLE management. Oleocanthal (OLE), the main extra virgin olive oil (EVOO)-derived secoiridoid, has shown to regulate the immune-inflammatory response in various disease contexts; however, its possible beneficial effects on SLE remain unclear. This study sought to evaluate the effects of OLE enriched diet on renal damage and aortic endothelial dysfunction in murine pristane-induced SLE, focusing on the action mechanisms and signaling pathways involved. BALB/c mice were injected with pristane and fed with OLE supplemented diet (0.01 % (w/w)) for six months. Levels of cytokines were measured by ELISA in lipopolysaccharide (LPS)-stimulated peritoneal macrophages and splenocytes. Presence of immunoglobulin G (IgG) and IgM immune complexes were examined by immunofluorescence and immunohistochemistry. Thoracic aortas were used to evaluate endothelial dysfunction. Western blotting was employed to detect signaling pathways and oxidative-inflammatory-related mediators. Dietary OLE supplementation reduced Th1/Th17 pro-inflammatory cytokines production and alleviated renal damage by decreasing immunoglobulin complexes deposition, and inflammation-mediating enzymes expression. The mechanisms underlying these protective effects could be related to the regulation of nuclear factor erythroid 2-related factor 2/Haem oxygenase 1 (Nrf-2/HO-1), mitogen-activated protein kinases (MAPKs), signal transducer and transcription activator of transcription (STAT-3), inflammasome and, nuclear factor kappa B (NF-κB) signaling pathways. Also, dietary OLE improved aortic endothelial dysfunction and vascular reactivity, normalizing endothelial nitric oxide synthase (eNOS) uncoupling, and NADPH oxidase-1 (NOX-1) overexpression. This study shows the immunomodulatory effects of OLE in an in vivo model of SLE by improving renal damage and regulating aortic endothelial dysfunction. These preliminary results provide OLE as a new therapeutic strategy in SLE management.

Redox-responsive polyurethane-polyurea nanoparticles targeting to aortic endothelium and atherosclerosis

Aortic endothelial cell dysfunction is an early trigger of atherosclerosis, the major cause of the cardiovascular disease (CVD). Nanomedicines targeting vascular endothelium and lesions hold great promise as therapeutic solutions to vascular disorders. This study investigates the vascular delivery efficacy of polyurethane-polyurea nanocapsules (Puua-NCs) with pH-synchronized shell cationization and redox-triggered release. Fluorescent lipophilic dye DiI was encapsulated into Puua-NCs of variable sizes and concentrations. Invitro cellular uptake studies with human aortic endothelial cells showed that these Puua-NCs were taken up by cells in a dose-dependent manner. In apolipoprotein E-deficient mice fed a Western diet, a model of atherosclerosis, circulating Puua-NCs were stable and accumulated in aortic endothelium and lesions within 24 hours after intravenous administration. Treatment with thiol-reducing and oxidizing reagents disrupted the disulfide bonds on the surface of internalized NCs, triggering disassembly and intracellular cargo release. Ultimately, Puua-NCs are a potential redox-controllable cardiovascular drug delivery system.

Abstract P128: Sexual Dimorphisms In Perivascular Adipose Tissue In A Novel Mouse Model Of Diet-Induced Metabolic Syndrome.

Metabolic syndrome (MetS) is a complex multifactorial disease that increases cardiovascular risk. Vascular dysfunction associated with MetS is strongly influenced by perivascular adipose tissue (PVAT) which acts as a reservoir for immune cells. Sexual dimorphisms of PVAT pathophysiology are poorly understood due to a lack of clinically relevant animal models of MetS that show metabolic disturbances in males and females. Using a new diet-induced model of MetS pathophysiology in both sexes, we characterised the aortic leukocyte profile and determined PVAT influence on vascular function. Six-week-old male and female C57BL/6 mice were fed either a high-fat diet (43% kcal) with high sugar and salt (10% high fructose and 0.9% NaCl; HFSS) in drinking water, or normal control diet (NCD) for 10 weeks. Physiological parameters were measured fortnightly. At endpoint, aorta with PVAT intact was isolated for leukocyte enumeration (flow cytometry) or for in vitro vascular reactivity (± PVAT; wire myography). Weight gain (NCD M: 29.3±0.5 g vs. MetS M: 37.5±1.0 g; NCD F: 23.6±0.7 g vs. MetS F: 28.9±1.2 g), fasting blood glucose (NCD M: 9.5±0.4 mmol/L vs. MetS M: 11.3 mmol/L; NCD F: 8.9±0.3 mmol/L vs. 10.4±0.5 mmol/L), blood cholesterol (NCD M: 125.0±3.0 mg/dL vs. MetS M: 177.5±6.5 mg/dL; NCD F: 96.7±2.9 mg/dL vs. MetS F: 130.7±3.4 mg/dL) and systolic blood pressure (NCD M: 121.0±1.8 mmHg vs. MetS M: 132.0±2.1 mmHg; NCD F: 122.3±2.2 mmHg vs. MetS F: 133.6±2.7 mmHg) were increased in both sexes of HFSS-fed mice (P<0.05 n= 12; vs. NCD) confirming that the HFSS model is a clinically relevant diet-induced model of MetS. Despite comparable metabolic disturbances in both sexes, aortic inflammation was only observed in males (increased pro-inflammatory monocytes (Ly6C hi+ ; M: 3.3±0.5 x 10 3 cells/aorta vs F: 1.6±0.4 x 10 3 cells/aorta) and neutrophils (Ly6G + ; M: 0.9±-0.2 x 10 3 cells/aorta vs F: 0.3+/0.1 x 10 3 cells/aorta); P<0.05 vs. HFSS-fed female. MetS altered PVAT and vascular function in both sexes, but with contrasting effects. PVAT blunted endothelium-dependent relaxation in HFSS-fed males (P<0.05 vs. NCD male) and in NCD-fed females (P<0.05 vs. HFSS females). The data indicates that sexual dimorphisms in PVAT impacts MetS-induced aortic inflammation and endothelial dysfunction.

Abstract P081: Early Life Stress (ELS) Induces Greater Aortic Pulse Wave Velocity (PWV) Through Reactive Oxygen Species (ROS)

ELS, such as verbal and physical abuse or maltreatment, is linked to a greater risk ofcardiovascular disease (CVD) in adulthood. Aortic stiffness, an independent predictor of CVD,has been suggested as a prognostic tool for early detection and intervention in at-riskpopulations. Several human studies have shown that aortic stiffness (assessed by PWV) isincreased in both adults and adolescents with ELS. ELS mouse models, such as maternalseparation with early weaning (MSEW), show aortic endothelial dysfunction with increasedsuperoxide and systemic inflammation, which are both present in humans with ELS exposureand contribute to aortic stiffness. We hypothesized that MSEW mice develop aortic stiffness in aROS-dependent manner at an early age. The MSEW protocol entails separating pups from thedam 4h/day (postnatal, PD 2 to 5) and 8h/day (PD 6 to 16) followed by weaning at PD17.Normally-reared (NR) litters remain with the dam and are weaned at PD21. Female and maleMSEW and NR mice were treated for 2 weeks with the ROS scavenger, TEMPOL (30mg/kg/day; drinking water) from 8 weeks of age. PWV was assessed prior to and after TEMPOLtreatment. PWV was significantly increased in MSEW mice at baseline (NR: 1.95 mm/sec ±0.15, MSEW: 2.60 mm/sec ± 0.17, p=0.009) in both sexes similarly. PWV was not significantlydifferent between MSEW and NR mice after TEMPOL treatment (NR: 1.78 mm/sec ± 0.06,MSEW: 1.97 mm/sec ± 0.12, p<0.05). MSEW mice had significantly more aorticCD11b + Ly6C + CX3CR1 + inflammatory monocytes compared to NR mice (NR: .9% ± 0.31,MSEW 1.5% ± 0.25, p=0.014) at baseline, however TEMPOL did not affect this. TEMPOLtreatment significantly decreased CD11b + F4/80 + MHCII hi inflammatory resident macrophages inMSEW mice compared to NR mice (Median Fluorescent Intensity; NR: 1,235,274 ± 54413,MSEW : 944,446 ±11728, p=0.007). In conclusion, ELS induces increased aortic stiffness in aROS-specific manner at an early age. Inflammatory resident macrophages may contribute to theELS-specific ROS-dependent mechanism. The mechanisms identified in this work arepotentially powerful tools for preventative therapeutics to reduce CVD burden in ELS-populations.

Contribution of Oxidative Stress (OS) in Calcific Aortic Valve Disease (CAVD): From Pathophysiology to Therapeutic Targets.

Calcific aortic valve disease (CAVD) is a major cause of cardiovascular mortality and morbidity, with increased prevalence and incidence. The underlying mechanisms behind CAVD are complex, and are mainly illustrated by inflammation, mechanical stress (which induces prolonged aortic valve endothelial dysfunction), increased oxidative stress (OS) (which trigger fibrosis), and calcification of valve leaflets. To date, besides aortic valve replacement, there are no specific pharmacological treatments for CAVD. In this review, we describe the mechanisms behind aortic valvular disease, the involvement of OS as a fundamental element in disease progression with predilection in AS, and its two most frequent etiologies (calcific aortic valve disease and bicuspid aortic valve); moreover, we highlight the potential of OS as a future therapeutic target.

CXCR6 Mediates Pressure Overload-Induced Aortic Stiffness by Increasing Macrophage Recruitment and Reducing Exosome-miRNA29b.

Aortic stiffness is an independent risk factor for aortic diseases such as aortic dissection which commonly occurred with aging and hypertension. Chemokine receptor CXCR6 is critically involved in vascular inflammation and remodeling. Here, we investigated whether and how CXCR6 plays a role in aortic stiffness caused by pressure overload. CXCR6-/- and WT mice underwent transverse aortic constriction (TAC) surgery for 8weeks. CXCR6 deficiency significantly improved TAC-induced aortic remodeling and endothelial dysfunction by decreasing CD11c+ macrophage infiltration, suppressing VCAM-1 and ICAM-1, reducing collagen deposition, and downregulating MMP12 and osteopontin in the aorta. Consistently, blocking the CXCL16/CXCR6 axis also reduced aortic accumulation of CD11c+ macrophages and vascular stiffness but without affecting the release of TNF-α and IL-6 from the aorta. Furthermore, pressure overload inhibited aortic release of exosomes, which could be reversed by suppressing CXCR6 or CXCL16. Inhibition of exosome release by GW4869 significantly aggravated TAC-induced aortic calcification and stiffness. By exosomal microRNA microarray analysis, we found that microRNA-29b was significantly reduced in aortic endothelial cells (AECs) receiving TAC. Intriguingly, blocking the CXCL16/CXCR6 axis restored the expression of miR-29b in AECs. Finally, overexpression of miR-29b significantly increased eNOS and reduced MMPs and collagen in AECs. By contrast, antagonizing miR-29b in vivo further enhanced TAC-induced expressions of MMP12 and osteopontin, aggravated aortic fibrosis, calcification, and stiffness. Our study demonstrated a key role of the CXCL16/CXCR6 axis in macrophage recruitment and macrophage-mediated aortic stiffness under pressure overload through an exosome-miRNAs-dependent manner.

Abstract P3013: MicroRNA-122 Targeting With Gamma Peptide Nucleic Acid Inhibitor Improves Vascular Endothelial Dysfunction In Pre-diabetic Mice.

Background: In patients with diabetes, the endothelial function precedes and predicts the risk of cardiovascular disorders. Current treatment approaches fail to decrease unwanted cardiovascular outcomes in these patients. The blood levels of microRNA-122 (miR-122) elevated in patients with diabetes and correlated with risk and severity of cardiovascular disorders. Therefore, targeting miR-122 with efficient and safer inhibitors could be a promising approach. The objective of this study was to investigate the effectiveness of gamma peptide nucleic acid (γPNA)-based miR-122 inhibitor in preventing the development of endothelial dysfunction in pre-diabetic mice. Methods: To test the effectiveness of γPNA technology in inhibiting miR-122, we synthesized γPNA-miR-122 inhibitor (γP-122-I) and γPNA-scrambled control (γP-SC). The purity and molecular weight of γP-122-I and γP-SC were assessed by HPLC and MALDI spectrometry, respectively. The binding and affinity of γP-122-I were determined by gel-shift assay and thermal denaturation. The insulin-resistance (induced by high-fat-diet, HFD) mice were injected with γP-122-I (5 mg/kg/day, i.p., 6 weeks). miR-122 expression was measured by real-time PCR. The safety of γP-122-I was evaluated by biochemical and histological analysis of vital organs (liver & kidney). The vascular endothelial function was assessed by determining endothelium-dependent and independent relaxation of phenylephrine-induced precontracted aortic rings. Results were expressed as mean ± s.e.m. Results: The HPLC of γPNA oligomers show that both γP-122-I and γP-SC were >95% pure. The molecular weight of γP-122-I (m/z: 9227.4) and γP-SC (m/z: 9232.7) were confirmed by MALDI spectrometry. γP-122-I strongly binds with miR-122 and forms a stable complex compared to commercially available miR-122 inhibitor. A comprehensive safety analysis of γP-122-I in mice shows that neither it affected liver and kidney function nor their morphology during acute and chronic exposure, respectively. Next, HFD-fed mice show higher serum and aortic miR-122 levels, endothelial dysfunction, and inflammation, which were reversed by γP-122-I treatment. Conclusions: γP-122-I inhibits the development of impaired endothelial function in pre-diabetic mice without any evidence of toxicity

Influenza A virus elicits peri-vascular adipose tissue inflammation and vascular dysfunction of the aorta in pregnant mice.

Influenza A virus (IAV) infection during pregnancy initiates significant aortic endothelial and vascular smooth muscle dysfunction, with inflammation and T cell activation, but the details of the mechanism are yet to be clearly defined. Here we demonstrate that IAV disseminates preferentially into the perivascular adipose tissue (PVAT) of the aorta in mice. IAV mRNA levels in the PVAT increased at 1–3 days post infection (d.p.i) with the levels being ~4–8 fold higher compared with the vessel wall. IAV infection also increased Ly6Clow patrolling monocytes and Ly6Chigh pro-inflammatory monocytes in the vessel wall at 3 d.p.i., which was then followed by a greater homing of these monocytes into the PVAT at 6 d.p.i. The vascular immune phenotype was characteristic of a “vascular storm”- like response, with increases in neutrophils, pro-inflammatory cytokines and oxidative stress markers in the PVAT and arterial wall, which was associated with an impairment in endothelium-dependent relaxation to acetylcholine. IAV also triggered a PVAT compartmentalised elevation in CD4+ and CD8+ activated T cells. In conclusion, the PVAT of the aorta is a niche that supports IAV dissemination and a site for perpetuating a profound innate inflammatory and adaptive T cell response. The manifestation of this inflammatory response in the PVAT following IAV infection may be central to the genesis of cardiovascular complications arising during pregnancy.

Enduring Reactive Oxygen Species Emission Causes Aberrant Protein S-Glutathionylation Transitioning Human Aortic Valve Cells from a Sclerotic to a Stenotic Phenotype.

Aims: During calcific aortic valve stenosis (CAVS) progression, oxidative stress and endothelial dysfunction mark the initial pathogenic steps with a parallel dysregulation of the antioxidant systems. Here, we tested whether oxidation-induced protein S-glutathionylation (P-SSG) accounts for a phenotypic switch in human aortic valvular tissue, eventually leading to calcium deposition. Next, we tested whether countering this reactive oxygen species (ROS) surge would prevent these perturbations. Results: We employed state-of-the-art technologies, such as electron paramagnetic resonance (EPR), liquid chromatography-tandem mass spectrometry, imaging flow-cytometry, and live-cell imaging on human excised aortic valves and primary valve endothelial cells (VECs). We observed that a net rise in EPR-detected ROS emission marked the transition from fibrotic to calcific in human CAVS specimens, coupled to a progressive increment in P-SSG deposition. In human VECs (hVECs), treatment with 2-acetylamino-3-[4-(2-acetylamino-2-carboxyethylsulfanylthiocarbonylamino)phenylthiocarbamoylsulfanyl]propionic acid triggered highly oxidizing conditions prompting P-SSG accumulation, damaging mitochondria, and inducing endothelial nitric oxide synthase uncoupling. All the events conjured up in morphing these cells from their native endothelial phenotype into a damaged calcification-inducing one. As proof of principle, the use of the antioxidant N-acetyl-L-cysteine prevented these alterations. Innovation: Borne as a compensatory system to face excessive oxidative burden, with time, P-SSG contributes to the morphing of hVECs from their innate phenotype into a damaged one, paving the way to calcium deposition. Conclusion: Our data suggest that, in the human aortic valve, unremitted ROS emission along with a P-SSG build-up occurs and accounts, at least in part, for the morphological/functional changes leading to CAVS. Antioxid. Redox Signal. 37, 1051-1071.

Exercise Training Alleviates Alzheimer’s Disease‐Associated Aortic Endothelial Dysfunction

BackgroundsA strong association between cardiovascular disease and Alzheimer’s disease (AD) has been reported, but its possible mechanism has been unknown. We have tested whether aortic endothelial function in AD is impaired and investigated the protective effect of exercise training (EX) on endothelial function in aorta of AD mice.METHODSWild type (C57BL/6, WT) and AD (APP/PS1dE9, AD) mice underwent treadmill training (WT‐EX and AD‐EX) or remained sedentary for 12 weeks. The expression of total‐and phosphorylated‐endothelial nitric oxide synthase (t‐eNOS and p‐eNOS), beta‐amyloid, and superoxide dismutase 2 (SOD2) were quantified by Western blot analysis.RESULTSt‐eNOS and p‐eNOS expressions in AD were decreased compared to WT but EX increased both t‐eNOS and p‐eNOS in AD (P < 0.01 and P = 0.09). Beta‐amyloid levels in AD were higher compared to WT but EX decreased it (P < 0.05). SOD2 levels in AD were increased by EX (P < 0.05) whereas there was no change in WT.CONCLUSIONRegular aerobic exercise alleviates e‐NOS‐mediated endothelial dysfunction in aorta of AD mice possibly via reduced beta‐amyloid and increased SOD2 levels.

Sarco/Endoplasmic Reticulum Ca2+ ATPase 2 Activator Ameliorates Endothelial Dysfunction; Insulin Resistance in Diabetic Mice.

Background: Sarco/endoplasmic reticulum Ca2+-ATPase2 (SERCA2) is impaired in various organs in animal models of diabetes. The purpose of this study was to test the effects of an allosteric SERCA2 activator (CDN1163) on glucose intolerance, hepatosteatosis, skeletal muscle function, and endothelial dysfunction in diabetic (db/db) mice. Methods: Either CDN1163 or vehicle was injected intraperitoneally into 16-week-old male control and db/db mice for 5 consecutive days. Results: SERCA2 protein expression was decreased in the aorta of db/db mice. In isometric tension measurements of aortic rings from db/db mice treated with CDN1163, acetylcholine (ACh)-induced relaxation was improved. In vivo intraperitoneal administrations of CDN 1163 also increased ACh-induced relaxation. Moreover, CDN1163 significantly decreased blood glucose in db/db mice at 60 and 120 min during a glucose tolerance test; it also decreased serum insulin levels, hepatosteatosis, and oxygen consumption in skeletal muscle during the early period of exercise in db/db mice. Conclusions: CDN1163 directly improved aortic endothelial dysfunction in db/db mice. Moreover, CDN1163 improved hepatosteatosis, skeletal muscle function, and insulin resistance in db/db mice. The activation of SERCA2 might be a strategy for the all the tissue expressed SERCA2a improvement of endothelial dysfunction and the target for the organs related to insulin resistance.

Comprehensive Characterization of Arterial and Cardiac Function in Marfan Syndrome—Can Biomarkers Help Improve Outcome?

Background: Marfan Syndrome (MFS) has been associated with increased aortic stiffness and left ventricular dysfunction. The latter may be due to the underlying genotype and/or secondary to aortic stiffening (vascular-ventricular interaction). The aim of this study was to characterize arterial and cardiac function in MFS using a multimodal approach. Methods: Prospective observational study of MFS patients and healthy controls. Methods included echocardiography, ascending aortic distensibility, common carotid intima media thickness [cIMT], parameters of wave reflection, carotid-femoral pulse wave velocity [cfPWV]), reactive hyperemia index [RHI], and biomarker analysis (Olink, CVII panel). Results: We included 20 patients with MFS and 67 controls. Ascending aortic distensibility, cIMT and RHI were decreased, while all parameters of arterial wave reflection, stiffness and BNP levels were increased in the MFS group. Both systolic and diastolic function were impaired relative to controls. Within the MFS group, no significant correlation between arterial and cardiac function was identified. However, cfPWV correlated significantly with indexed left ventricular mass and volume in MFS. Bran natriuretic peptide (BNP) was the only biomarker significantly elevated in MFS following correction for age and sex. Conclusions: MFS patients have generally increased aortic stiffness, endothelial dysfunction and BNP levels while cIMT is decreased, supporting that the mechanism of general stiffening is different from acquired vascular disease. CfPWV is associated with cardiac size, blood pressure and BNP in MFS patients. These may be early markers of disease progression that are suitable for monitoring pharmacological treatment effects in MFS patients.

Abdominal Aortic Endothelial Dysfunction Occurs in Female Mice With Dextran Sodium Sulfate-Induced Chronic Colitis Independently of Reactive Oxygen Species Formation.

Background and ObjectiveInflammatory bowel disease (IBD) produces significant local and systemic inflammation with increased reactive oxygen species (ROS) formation. IBD Patients are at an increased risk for developing endothelial dysfunction and cardiovascular diseases. The present study tested the hypothesis that IBD impairs aortic endothelial function via ROS formation and investigate potential sex-related differences.Methods and ResultsAcute and chronic colitis models were induced in male and female C57BL/6 mice with dextran sodium sulfate (DSS) treatment. Aortic wall stiffness, endothelial function, and ROS levels, as well as serum levels of pro-inflammatory cytokines were evaluated. Acetylcholine (Ach)-induced endothelium-dependent relaxation of abdominal aorta without perivascular adipose tissue (PVAT) was significantly reduced in female mice, not males, with chronic colitis without a change in nitroglycerin-induced endothelium-independent relaxation. PVAT effectively preserved Ach-induced relaxation in abdominal aorta of female mice with chronic colitis. Aortic peak velocity, maximal intraluminal diameters, pulse wave velocity, distensibility and radial strain were preserved in mice with both acute and chronic colitis. Although pro-inflammatory cytokines levels were increased in mice with acute and chronic colitis, aortic ROS levels were not increased.ConclusionThe data demonstrate that abdominal aortic endothelial function was attenuated selectively in female mice with chronic colitis independent of ROS formation. Further, PVAT played an important role in preserving endothelial function in female mice with chronic colitis.

Silibinin augments the effect of clopidogrel on atherosclerosis in diabetic ApoE deficiency mice.

Diabetes mellitus (DM) abolishes the antithrombotic effect of Clopidogrel. Here, we investigated the synergistic effect of Silibinin on Clopidogrel-mediated atherosclerosis treatment in diabetic mice. ApoE-/- mice were fed with high-fat diet (HFD) to establish the atherosclerotic model with diabetes. Animals were treated with Clopidogrel, Silibinin, or the combined to evaluate the protective effects on atherosclerosis and diabetes through Oil-red-O staining, qRT-PCR, Western blot, and metabolic measurements. Platelet activation and aggregation ex vivo assays were performed to detect the anti-thrombotic effect of different administrations. Silibinin significantly enhanced the inhibitory effect of Clopidogrel on atherosclerosis in DM mice. Co-administration of Silibinin with Clopidogrel remarkedly reduced the aortic lesion, inflammation, and endothelial dysfunction in aorta roots, and diabetic symptoms were significantly improved by the Silibinin-Clopidogrel treatment in HFD-fed ApoE-/- mice. Interestingly, the anti-thrombotic effect of Clopidogrel was further augmented by the Silibinin treatment in atherosclerotic mice. In atherosclerotic mouse model, Silibinin significantly improves the effect of Clopidogrel on atherosclerosis.

CagA+Helicobacter pylori, Not CagA–Helicobacter pylori, Infection Impairs Endothelial Function Through Exosomes-Mediated ROS Formation

BackgroundHelicobacter pylori (H. pylori) infection increases the risk for atherosclerosis, and ROS are critical to endothelial dysfunction and atherosclerosis. CagA is a major H. pylori virulence factor associated with atherosclerosis. The present study aimed to test the hypothesis that CagA+ H. pylori effectively colonizes gastric mucosa, and CagA+ H. pylori, but not CagA– H. pylori, infection impairs endothelial function through exosomes-mediated ROS formation.MethodsC57BL/6 were used to determine the colonization ability of CagA+ H. pylori and CagA– H. pylori. ROS production, endothelial function of thoracic aorta and atherosclerosis were measured in CagA+ H. pylori and CagA– H. pylori infected mice. Exosomes from CagA+ H. pylori and CagA– H. pylori or without H. pylori infected mouse serum or GES-1 were isolated and co-cultured with bEND.3 and HUVECs to determine how CagA+ H. pylori infection impairs endothelial function. Further, GW4869 was used to determine if CagA+H. pylori infection could lead to endothelial dysfunction and atherosclerosis through an exosomes-mediated mechanism.ResultsCagA+ H. pylori colonized gastric mucosa more effectively than CagA– H. pylori in mice. CagA+ H. pylori, not CagA– H. pylori, infection significantly increased aortic ROS production, decreased ACh-induced aortic relaxation, and enhanced early atherosclerosis formation, which were prevented with N-acetylcysteine treatment. Treatment with CagA-containing exosomes significantly increased intracellular ROS production in endothelial cells and impaired their function. Inhibition of exosomes secretion with GW4869 effectively prevented excessive aortic ROS production, endothelial dysfunction, and atherosclerosis in mice with CagA+ H. pylori infection.ConclusionThese data suggest that CagA+ H. pylori effectively colonizes gastric mucosa, impairs endothelial function, and enhances atherosclerosis via exosomes-mediated ROS formation in mice.

SIRT3-AMPK signaling pathway as a protective target in endothelial dysfunction of early sepsis

Extensive vascular endothelial dysfunction usually occurs in sepsis, resulting in high mortality. The purpose of this study was therefore to investigate the role of AMP-dependent protein kinase (AMPK) in the aortic endothelial dysfunction of early sepsis in mice, and the relationship between AMPK and Sirtuin3 (SIRT3). Cecal ligation and puncture (CLP) surgery was performed to establish a mouse sepsis model, and human umbilical vein endothelial cells (HUVECs) were treated with lipopolysaccharide (LPS) to mimic a sepsis model in vitro. We suppressed and increased the activities of AMPK with Dorsomorphin (CC) and Acadesine (AICAR), respectively. 3-TYP (SIRT3 inhibitor) and Honokiol (SIRT3 agonist) were used to alter SIRT3 activity. Then, the inflammatory and endothelial function parameters of the vascular tissue and survival rate were determined. In vivo, the expression of Ser1177 phosphorylation of endothelial nitric oxide synthase (p-eNOS), endothelium-dependent relaxation function, and survival decreased (P < 0.05), while NF-κB and NLRP3 pathways were activated in CLP-induced early sepsis (P < 0.05). Moreover, activation of AMPK significantly reversed the reduction of p-eNOS expression (P < 0.05), prevented endothelial dysfunction (P < 0.05), deactivated NF-κB and NLRP3 pathways (P < 0.05), and improved survival (P < 0.05) in septic mice. However, AMPK inhibition led to opposite effects (P < 0.05). In addition, changing the activity of AMPK had little effect on SIRT3 expression (P > 0.05), while the expression of p-AMPK varied with the inhibition or activation of SIRT3 (P < 0.05), which was further demonstrated using in vitro experiments. Together, the results showed that the SIRT3-AMPK signaling pathway played an important role in inhibiting vascular inflammation and endothelial dysfunction during early sepsis.

Lipoprotein(a), a Lethal Player in Calcific Aortic Valve Disease.

Calcified aortic valve disease (CAVD) is the most common valvular cardiovascular disease with increasing incidence and mortality. The primary treatment for CAVD is surgical or transcatheter aortic valve replacement and there remains a lack of effective drug treatment. Recently, lipoprotein (a) (Lp(a)) has been considered to play a crucial role in CAVD pathophysiology. Multiple studies have shown that Lp(a) represents an independent risk factor for CAVD. Moreover, Lp(a) mediates the occurrence and development of CAVD by affecting aortic valve endothelial dysfunction, indirectly promoting foam cell formation through oxidized phospholipids (OxPL), inflammation, oxidative stress, and directly promotes valve calcification. However, there is a lack of clinical trials with Lp(a) reduction as a primary endpoint. This review aims to explore the relationship and mechanism between Lp(a) and CAVD, and focuses on the current drugs that can be used as potential therapeutic targets for CAVD.

Repeated glucose spikes and insulin resistance synergistically deteriorate endothelial function and bardoxolone methyl ameliorates endothelial dysfunction.

BackgroundBoth insulin resistance and postprandial glucose spikes are known for their potential to induce vascular endothelial dysfunction in individuals with metabolic syndrome. However, these factors are inextricable, and therefore, their relative contributions to inducing endothelial dysfunction remain elusive. In this study, we aimed to disentangle the effects of these factors and clarify whether bardoxolone methyl (CDDO-Me), a novel nuclear factor erythroid 2-related factor 2 (Nrf2) activator, protects against glucose spike-induced endothelial dysfunction.MethodsWe induced glucose spikes twice daily for a duration of 1 week to rats fed a standard/control diet (CD) and Western-type diet (WTD). Endothelium-dependent relaxation (EDR) was evaluated using isolated thoracic aortas. Gene expression and dihydroethidium (DHE)-fluorescence studies were carried out; the effect of CDDO-Me on aortic endothelial dysfunction in vivo was also evaluated.ResultsNeither WTD-induced insulin resistance nor pure glucose spikes significantly deteriorated EDR. However, under high-glucose (20 mM) conditions, the EDR of thoracic aortas of WTD-fed rats subjected to glucose spikes was significantly impaired. In this group of rats, we observed significantly enhanced DHE fluorescence as a marker of reactive oxygen species, upregulation of an oxidative stress-related gene (NOX2), and downregulation of an antioxidant gene (SOD2) in the thoracic aortas. As expected, treatment of the thoracic aorta of this group of rats with antioxidant agents significantly improved EDR. We also noted that pretreatment of aortas from the same group with CDDO-Me attenuated endothelial dysfunction, accompanied by a correction of the redox imbalance, as observed in gene expression and DHE fluorescence studies.ConclusionsFor the first time, we showed that insulin resistance and glucose spikes exert a synergistic effect on aortic endothelial dysfunction. Furthermore, our study reveals that CDDO-Me ameliorates endothelial dysfunction caused by glucose spikes in a rat model of metabolic syndrome.