Cannabinoid Receptor 2 Signaling Does Not Modulate Atherogenesis in Mice

BACKGROUND:Strong evidence supports a protective role of the cannabinoid receptor 2 (CB(2)) in inflammation and atherosclerosis. However, direct proof of its involvement in lesion formation is lacking. Therefore, the present study aimed to characterize the role of the CB(2) receptor in Murine atherogenesis. METHODS AND FINDINGS:Low density lipoprotein receptor-deficient (LDLR(-/-)) mice subjected to intraperitoneal injections of the selective CB(2) receptor agonist JWH-133 or vehicle three times per week consumed high cholesterol diet (HCD) for 16 weeks. Surprisingly, intimal lesion size did not differ between both groups in sections of the aortic roots and arches, suggesting that CB(2) activation does not modulate atherogenesis in vivo. Plaque content of lipids, macrophages, smooth muscle cells, T cells, and collagen were also similar between both groups. Moreover, CB(2) (-/-)/LDLR(-/-) mice developed lesions of similar size containing more macrophages and lipids but similar amounts of smooth muscle cells and collagen fibers compared with CB(2) (+/+)/LDLR(-/-) controls. While JWH-133 treatment reduced intraperitoneal macrophage accumulation in thioglycollate-elicited peritonitis, neither genetic deficiency nor pharmacologic activation of the CB(2) receptor altered inflammatory cytokine expression in vivo or inflammatory cell adhesion in the flow chamber in vitro. CONCLUSION:Our study demonstrates that both activation and deletion of the CB(2) receptor do not relevantly modulate atherogenesis in mice. Our data do not challenge the multiple reports involving CB(2) in other inflammatory processes. However, in the context of atherosclerosis, CB(2) does not appear to be a suitable therapeutic target for reduction of the atherosclerotic plaque

CD40L Deficiency Attenuates Diet-Induced Adipose Tissue Inflammation by Impairing Immune Cell Accumulation and Production of Pathogenic IgG-Antibodies

BACKGROUND: Adipose tissue inflammation fuels the metabolic syndrome. We recently reported that CD40L--an established marker and mediator of cardiovascular disease--induces inflammatory cytokine production in adipose cells in vitro. Here, we tested the hypothesis that CD40L deficiency modulates adipose tissue inflammation in vivo. METHODOLOGY/PRINCIPAL FINDINGS: WT or CD40L(-/-) mice consumed a high fat diet (HFD) for 20 weeks. Inflammatory cell recruitment was impaired in mice lacking CD40L as shown by a decrease of adipose tissue macrophages, B-cells, and an increase in protective T-regulatory cells. Mechanistically, CD40L-deficient mice expressed significantly lower levels of the pro-inflammatory chemokine MCP-1 both, locally in adipose tissue and systemically in plasma. Moreover, levels of pro-inflammatory IgG-antibodies against oxidized lipids were reduced in CD40L(-/-) mice. Also, circulating low-density lipoproteins and insulin levels were lower in CD40L(-/-) mice. However, CD40L(-/-) mice consuming HFD were not protected from the onset of diet-induced obesity (DIO), insulin resistance, and hepatic steatosis, suggesting that CD40L selectively limits the inflammatory features of diet-induced obesity rather than its metabolic phenotype. Interestingly, CD40L(-/-) mice consuming a low fat diet (LFD) showed both, a favorable inflammatory and metabolic phenotype characterized by diminished weight gain, improved insulin tolerance, and attenuated plasma adipokine levels. CONCLUSION: We present the novel finding that CD40L deficiency limits adipose tissue inflammation in vivo. These findings identify CD40L as a potential mediator at the interface of cardiovascular and metabolic disease

Augmenting SOD2 Levels in Apoe-/- Mice with Peptide-mRNA Nanoparticles Preserves Atherosclerotic Plaque Stability in Advanced Atherosclerosis

http://deepblue.lib.umich.edu/bitstream/2027.42/191312/2/Sod2-p5RHH in athero poster 2023.pd

Mitochondrial Oxidative Stress Mediates Macrophage Pro-inflammatory Metabolic Switch in Atherosclerotic Vascular Disease in Aging

Aging elevates cardiovascular disease risk, including atherosclerosis. Macrophages play crucial role in vascular aging by promoting inflammation and atherosclerosis progression. Age-related increase in NOX4 NADPH oxidase expression correlates with mitochondrial dysfunction, inflammation, and atherosclerosis severity. We hypothesized that NOX4-dependent mitochondrial oxidative stress induces macrophage metabolic dysfunction and an inflammatory phenotype in aging-associated atherosclerotic disease. Aortic and brachiocephalic artery lesion areas were comparable in 5-month-old (young) Nox4-/-/Apoe-/- and Apoe-/- mice, increased significantly in 16-month-old (aged) mice, but were significantly lower in Nox4-/-/Apoe-/- versus Apoe-/- mice. In aged Nox4-/-/Apoe-/- mice, atherosclerotic lesions had reduced CD11b+ area, lower expression of CCL2, IL1β, and IL6, and fewer classically activated pro-inflammatory macrophages (CD38+CD80+). Notably, there was also an increased proportion of alternatively activated pro-resolving macrophages (CD163+CD206+). Spectral flow cytometry and t-SNE analysis revealed a significantly lower proportion of activated inflammatory macrophages and macrophage-like cells in atherosclerotic lesions of aged Nox4-/-/Apoe-/- compared to Apoe-/- mice. Macrophages from aged Apoe-/- mice had altered metabolic function. In contrast, macrophages from Nox4-/-/Apoe-/- mice were less glycolytic, more aerobic, and had preserved basal and maximal respiration and mitochondrial ATP production. Nox4-/-/Apoe-/- macrophages had lower mitochondrial ROS and reduced IL1β secretion, compared with Apoe-/- mice. In aged Apoe-/- mice, inhibition of NOX4 using GKT137831 significantly reduced macrophage ROS and improved mitochondrial function. This resulted in a decreased CD68+CD80+ and increased CD163+CD206+ lesion macrophages and attenuated atherosclerosis. Our results imply that NOX4-dependent mitochondrial oxidative stress in aging contributes to macrophage mitochondrial dysfunction, glycolytic metabolic switch, and pro-inflammatory phenotype, advancing atherosclerosis. Inhibition of NOX4 could alleviate vascular inflammation and atherosclerosis by improving mitochondrial function in macrophages.http://deepblue.lib.umich.edu/bitstream/2027.42/191311/2/AVendrov poster 2023 R.pd

NOXA1-dependent NADPH oxidase regulates redox signaling and phenotype of vascular smooth muscle cell during atherogenesis

Increased reactive oxygen species (ROS) production and inflammation are key factors in the pathogenesis of atherosclerosis. We previously reported that NOX activator 1 (NOXA1) is the critical functional homolog of p67phox for NADPH oxidase activation in mouse vascular smooth muscle cells (VSMC). Here we investigated the effects of systemic and SMC-specific deletion of Noxa1 on VSMC phenotype during atherogenesis in mice.Neointimal hyperplasia following endovascular injury was lower in Noxa1-deficient mice versus the wild-type following endovascular injury. Noxa1 deletion in Apoe-/- or Ldlr-/- mice fed a Western diet showed 50% reduction in vascular ROS and 30% reduction in aortic atherosclerotic lesion area and aortic sinus lesion volume (P < 0.01). SMC-specific deletion of Noxa1 in Apoe-/- mice (Noxa1SMC-/-/Apoe-/-) similarly decreased vascular ROS levels and atherosclerotic lesion size. TNFα-induced ROS generation, proliferation and migration were significantly attenuated in Noxa1-deficient versus wild-type VSMC. Immunofluorescence analysis of atherosclerotic lesions showed a significant decrease in cells positive for CD68 and myosin11 (22% versus 9%) and Mac3 and α-actin (17% versus 5%) in the Noxa1SMC-/-/Apoe-/- versus Apoe-/- mice. The expression of transcription factor KLF4, a modulator of VSMC phenotype, and its downstream targets – VCAM1, CCL2, and MMP2 – were significantly reduced in the lesions of Noxa1SMC-/-/Apoe-/- versus Apoe-/- mice as well as in oxidized phospholipids treated Noxa1SMC-/- versus wild-type VSMC.Our data support an important role for NOXA1-dependent NADPH oxidase activity in VSMC plasticity during restenosis and atherosclerosis, augmenting VSMC proliferation and migration and KLF4-mediated transition to macrophage-like cells, plaque inflammation, and expansion. Keywords: Oxidative stress, NOXA1, Smooth muscle cells, KLF4, Macrophage-like cells, Atherosclerosi

NADPH Oxidases and Oxidative Stress in the Pathogenesis of Atrial Fibrillation.

Atrial fibrillation (AF) is the most common type of cardiac arrhythmia and its prevalence increases with age. The irregular and rapid contraction of the atria can lead to ineffective blood pumping, local blood stasis, blood clots, ischemic stroke, and heart failure. NADPH oxidases (NOX) and mitochondria are the main sources of reactive oxygen species in the heart, and dysregulated activation of NOX and mitochondrial dysfunction are associated with AF pathogenesis. NOX- and mitochondria-derived oxidative stress contribute to the onset of paroxysmal AF by inducing electrophysiological changes in atrial myocytes and structural remodeling in the atria. Because high atrial activity causes cardiac myocytes to expend extremely high energy to maintain excitation-contraction coupling during persistent AF, mitochondria, the primary energy source, undergo metabolic stress, affecting their morphology, Ca2+ handling, and ATP generation. In this review, we discuss the role of oxidative stress in activating AF-triggered activities, regulating intracellular Ca2+ handling, and functional and anatomical reentry mechanisms, all of which are associated with AF initiation, perpetuation, and progression. Changes in the extracellular matrix, inflammation, ion channel expression and function, myofibril structure, and mitochondrial function occur during the early transitional stages of AF, opening a window of opportunity to target NOX and mitochondria-derived oxidative stress using isoform-specific NOX inhibitors and mitochondrial ROS scavengers, as well as drugs that improve mitochondrial dynamics and metabolism to treat persistent AF and its transition to permanent AF.http://deepblue.lib.umich.edu/bitstream/2027.42/191308/2/NADPH Oxidases and Oxidative Stress in the Pathogenesis of Atrial Fibrillation.pdfPublished onlineDescription of NADPH Oxidases and Oxidative Stress in the Pathogenesis of Atrial Fibrillation.pdf : Published versio

Bacterial Communities Associated with Atherosclerotic Plaques from Russian Individuals with Atherosclerosis.

Atherosclerosis is considered a chronic disease of the arterial wall and is the major cause of severe disease and death among individuals all over the world. Some recent studies have established the presence of bacteria in atherosclerotic plaque samples and suggested their possible contribution to the development of cardiovascular disease. The main objective of this preliminary pilot study was to better understand the bacterial diversity and abundance in human atherosclerotic plaques derived from common carotid arteries of individuals with atherosclerosis (Russian nationwide group) and contribute towards the further identification of a main group of atherosclerotic plaque bacteria by 454 pyrosequencing their 16S ribosomal RNA (16S rRNA) genes. The applied approach enabled the detection of bacterial DNA in all atherosclerotic plaques. We found that distinct members of the order Burkholderiales were present at high levels in all atherosclerotic plaques obtained from patients with atherosclerosis with the genus Curvibacter being predominant in all plaque samples. Moreover, unclassified Burkholderiales as well as members of the genera Propionibacterium and Ralstonia were typically the most significant taxa for all atherosclerotic plaques. Other genera such as Burkholderia, Corynebacterium and Sediminibacterium as well as unclassified Comamonadaceae, Oxalobacteraceae, Rhodospirillaceae, Bradyrhizobiaceae and Burkholderiaceae were always found but at low relative abundances of the total 16S rRNA gene population derived from all samples. Also, we found that some bacteria found in plaque samples correlated with some clinical parameters, including total cholesterol, alanine aminotransferase and fibrinogen levels. Finally, our study indicates that some bacterial agents at least partially may be involved in affecting the development of cardiovascular disease through different mechanisms

Mitochondrial DAMPs-dependent inflammasome activation during aging induces vascular smooth muscle cell dysfunction and aortic stiffness in low aerobic capacity rats

Introduction: Low aerobic exercise capacity is an independent risk factor for cardiovascular disease (CVD) and a predictor of premature death. In combination with aging, low aerobic capacity lowers the threshold for CVD. Aim: Since low aerobic capacity and aging have been linked to mitochondrial oxidative stress and dysfunction, we investigated whether aged Low-Capacity Runner (LCR) rats (27 months) had vascular dysfunction compared to High-Capacity Runner (HCR) rats. Methods and Results: A significant decrease in aortic eNOS levels and vasodilation as well as an increase in aortic collagen and stiffness were observed in aged LCR rats compared to age and sex-matched HCR rats. There was a correlation between age-related vascular dysfunction and increased levels of ROS and DNA damage in aortas of LCR rats. Moreover, mitochondrial oxygen consumption, membrane potential, ATP levels, and mitophagy were lower in VSMCs of aged LCR rats. VSMCs from older LCR rats showed AIM2 inflammasome activation. VSMCs of young (4 months old) LCR rats treated with purified mitochondrial damage-associated molecular patterns (DAMP) recapitulated an inflammasome activation phenotype similar to that seen in aged rat VSMCs. Rapamycin, a potent immunosuppressant, induced mitophagy, stimulated electron transport chain activity, reduced inflammasome activity, mitochondrial ROS and DAMP levels in VSMCs from aged LCR rats. MitoTEMPO, a mitochondrial ROS scavenger, was similarly effective on VSMCs from aged rats.Conclusion: The findings suggest that impaired mitophagy and inflammasome activation in the vasculature under conditions of low aerobic exercise capacity during aging results in arterial dysfunction and aortic stiffness. In older adults with reduced aerobic capacity, mitochondrial antioxidants, mitophagy induction, and inflammasome inhibition may be effective therapeutic strategies for enhancing vascular health

Mitochondrial DAMPs-dependent inflammasome activation during aging induces vascular smooth muscle cell dysfunction and aortic stiffness in low aerobic capacity rats

Introduction: Low aerobic exercise capacity is an independent risk factor for cardiovascular disease (CVD) and a predictor of premature death. In combination with aging, low aerobic capacity lowers the threshold for CVD. Aim: Since low aerobic capacity and aging have been linked to mitochondrial oxidative stress and dysfunction, we investigated whether aged Low-Capacity Runner (LCR) rats (27 months) had vascular dysfunction compared to High-Capacity Runner (HCR) rats. Methods and Results: A significant decrease in aortic eNOS levels and vasodilation as well as an increase in aortic collagen and stiffness were observed in aged LCR rats compared to age and sex-matched HCR rats. There was a correlation between age-related vascular dysfunction and increased levels of ROS and DNA damage in aortas of LCR rats. Moreover, mitochondrial oxygen consumption, membrane potential, ATP levels, and mitophagy were lower in VSMCs of aged LCR rats. VSMCs from older LCR rats showed AIM2 inflammasome activation. VSMCs of young (4 months old) LCR rats treated with purified mitochondrial damage-associated molecular patterns (DAMP) recapitulated an inflammasome activation phenotype similar to that seen in aged rat VSMCs. Rapamycin, a potent immunosuppressant, induced mitophagy, stimulated electron transport chain activity, reduced inflammasome activity, mitochondrial ROS and DAMP levels in VSMCs from aged LCR rats. MitoTEMPO, a mitochondrial ROS scavenger, was similarly effective on VSMCs from aged rats. Conclusion: The findings suggest that impaired mitophagy and inflammasome activation in the vasculature under conditions of low aerobic exercise capacity during aging results in arterial dysfunction and aortic stiffness. In older adults with reduced aerobic capacity, mitochondrial antioxidants, mitophagy induction, and inflammasome inhibition may be effective therapeutic strategies for enhancing vascular health.http://deepblue.lib.umich.edu/bitstream/2027.42/175280/2/Mitochondrial DAMPs-dependent inflammasome activation during aging induces vascular smooth muscle cell dysfunction and aortic stiffness in low aerobic capacity rats_Canugovi-JCA 2022.pdfPublished onlineDescription of Mitochondrial DAMPs-dependent inflammasome activation during aging induces vascular smooth muscle cell dysfunction and aortic stiffness in low aerobic capacity rats_Canugovi-JCA 2022.pdf : Published versio

Vegetation and climate history in the Laptev Sea region (Arctic Siberia) during Late Quaternary inferred from pollen records

Paleoenvironmental records from a number of permafrost sections and lacustrine cores from the Laptev Sea region dated by several methods (14C-AMS, TL, IRSL, OSL and 230Th/U) were analyzed for pollen and palynomorphs. The records reveal the environmental history for the last ca 200 kyr. For interglacial pollen spectra, quantitative temperature values were estimated using the best modern analogue method. Sparse grass-sedge vegetation indicating arctic desert environmental conditions existed prior to 200 kyr ago. Dense, wet grass-sedge tundra habitats dominated during an interstadial ca 200-190 kyr ago, reflecting warmer and wetter summers than before. Sparser vegetation communities point to much more severe stadial conditions ca 190-130 kyr ago. Open grass and Artemisia communities with shrub stands (Alnus fruticosa, Salix, Betula nana) in more protected and moister places characterized the beginning of the Last Interglacial indicate climate conditions similar to present. Shrub tundra (Alnus fruticosa and Betula nana) dominated during the middle Eemian climatic optimum, when summer temperatures were 4-5 °C higher than today. Early-Weichselian sparse grass-sedge dominated vegetation indicates climate conditions colder and dryer than in the previous interval. Middle Weichselian Interstadial records indicate moister and warmer climate conditions, for example, in the interval 40-32 kyr BP Salix was present within dense, grass-sedge dominated vegetation. Sedge-grass-Artemisia-communities indicate that climate became cooler and drier after 30 kyr BP, and cold, dry conditions characterized the Late Weichselian, ca 26-16 kyr BP, when grass-dominated communities with Caryophyllaceae, Asteraceae, Cichoriaceae, Selaginella rupestris were present. From 16 to 12 kyr BP, grass-sedge communities with Caryophyllaceae, Asteraceae, and Cichoriaceae indicate climate was significantly warmer and moister than during the previous interval. The presence of Salix and Betula reflect temperatures about 4 °C higher than present at about 12-11 kyr BP, during the Allerød interval, but shrubs were absent in the Younger Dryas interval, pointing to a deterioration of climate conditions. Alnus fruticosa, Betula nana, Poaceae, and Cyperaceae dominate early Holocene spectra. Reconstructed absolute temperature values were substantially warmer than present (up to 12 °C). Shrubs gradually disappeared from coastal areas after 7.6 kyr BP when vegetation cover became similar to modern. A comparison of proxy-based paleoenvironmental reconstructions with the simulations performed by an Earth system model of intermediate complexity (CLIMBER-2) show good accordance between the regional paleodata and model simulations, especially for the warmer intervals. © 2011 Elsevier Ltd