Morelloflavone, a biflavonoid inhibitor of migration-related kinases, ameliorates atherosclerosis in mice

While macrophages take up modified LDL to form foam cells and multiply to develop fatty streaks, vascular smooth muscle cells (VSMC) migrate from the media to intima, secrete extracellular matrix, and increase the volume of atherosclerotic lesions. A medicinal plant Garcinia dulcis has been used in traditional Thai medicine for centuries to treat various chronic human diseases. Morelloflavone, a biflavonoid and an active ingredient of the plant, has been shown to inhibit VSMC migration through its inhibition of multiple migration-related kinases such as focal adhesion kinase, c-Src, ERK, and RhoA. However, the exact role of morelloflavone in atherosclerogenesis was unknown. We fed Ldlr−/−Apobec1−/− mice with either normal chow or chow containing 0.003% morelloflavone for 8 mo and assessed the extent of atherosclerosis by the en face and cross-sectional analyses. A cell composition analysis of atherosclerotic tissue was carried out using immunohistochemical staining. Oral morelloflavone therapy significantly reduced the atherosclerotic areas of the mouse aortas (a 26% reduction), without changing plasma lipid profiles or weights. Immunohistochemical analyses showed that morelloflavone reduced the number of VSMC in the atherosclerotic lesion while it did not change the density of macrophages in the lesion or the percentages of proliferating and apoptotic cells. Oral, low-dose, morelloflavone therapy retards atherosclerogenesis by limiting the migration of VSMC into the intima in the mouse model of human atherosclerosis. Upon further investigation, morelloflavone may be found to be a novel oral antiatherosclerotic agent and a viable addition to the conventional therapies such as statins in humans.

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Abstract 643: Type-2 Cannabinoid Receptor Deficiency is Associated with Atherosclerotic Lesion Calcification in Ldlr-null Mice

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.36.suppl_1.643 ◽

2016 ◽

Vol 36 (suppl_1) ◽

Author(s):

Makenzie L Fulmer ◽

Emilee Englehaupt ◽

Chris Garst ◽

Stacy Brown ◽

Douglas Thewke

Keyword(s):

Transcription Factor ◽

Cannabinoid Receptor ◽

Atherosclerotic Lesion ◽

Increased Risk ◽

Calcified Plaque ◽

En Face ◽

Related Transcription Factor ◽

A Cell ◽

Complex Signaling

Background: Calcification of atherosclerotic plaques is associated with vulnerability to rupture and increased risk of myocardial infarction. The mechanism of plaque calcification is unclear, but has been shown to be a cell-mediated process involving complex signaling pathways affecting the osteogenic transcription factor Runt-related transcription factor 2 (Runx2). The type-2 cannabinoid receptor (CB2) modulates processes involved in bone remodeling and our prior studies determined that CB2 alters the composition of early lesions in hyperlipidemic Ldlr -/- mice; however, the function of CB2 in plaque calcification is unknown. Therefore, we tested the hypothesis that CB2 modulates plaque calcification by evaluating the effects of systemic CB2 gene deletion on lesion calcification and aortic expression of Runx2 in Ldlr -/- mice. Results: Groups (n≥8) of 8-week old CB2 +/+ Ldlr -/- (WT) and CB2 -/- Ldlr -/- (CB2 -/- ) mice were fed a high fat diet (HFD) for up to 24 weeks. Standard blood plasma analysis showed no difference in HFD-induced hyperlipidemia between WT and CB2 -/- mice. Aortic levels of endocannabinoids, anandamide and 2-archidonylglycerol, were significantly elevated after 12 weeks of HFD feeding as determined by LC-MS/MS. En face analysis revealed the extent of atherosclerosis in the aortic arch and thoracic aorta did not differ between WT and CB2 -/- mice, but was ~1.9-fold greater in the abdominal aortas of CB2 -/- mice (17.0±1.3% vs 9.0±1.3%, p=0.002). Calcification of aortic root lesions was ~2.3 fold greater in CB2 -/- mice compared to WT mice (12.9±1.1% vs 5.6±1.2%, p=0.002) as revealed by von Kossa staining. Western blot analysis showed significantly increased expression of Runx2 in aortas of WT mice compared to CB2 -/- after 20 weeks of HFD (2.55±0.25 fold, p<0.05). Conclusion: Systemic CB2 deficiency enhances lesion calcification and is associated with altered aortic expression of Runx2. These results provide novel mechanistic insights into the function of CB2 signaling in the pathogenesis of atherosclerosis and vascular calcification that may lead to the development of therapies aimed at stabilizing calcified plaque.

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Flow-dependent YAP/TAZ activities regulate endothelial phenotypes and atherosclerosis

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1613121113 ◽

2016 ◽

Vol 113 (41) ◽

pp. 11525-11530 ◽

Cited By ~ 128

Author(s):

Kuei-Chun Wang ◽

Yi-Ting Yeh ◽

Phu Nguyen ◽

Elaine Limqueco ◽

Jocelyn Lopez ◽

...

Keyword(s):

Target Genes ◽

Critical Role ◽

Atherosclerotic Lesion ◽

Tissue Growth ◽

Binding Motif ◽

Disturbed Flow ◽

Atherosclerotic Lesions ◽

En Face

The focal nature of atherosclerotic lesions suggests an important role of local hemodynamic environment. Recent studies have demonstrated significant roles of Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) in mediating mechanotransduction and vascular homeostasis. The objective of this study is to investigate the functional role of YAP/TAZ in the flow regulation of atheroprone endothelial phenotypes and the consequential development of atherosclerotic lesions. We found that exposure of cultured endothelial cells (ECs) to the atheroprone disturbed flow resulted in YAP/TAZ activation and translocation into EC nucleus to up-regulate the target genes, including cysteine-rich angiogenic inducer 61 (CYR61), connective tissue growth factor (CTGF), and ankyrin repeat domain 1 (ANKRD1). In contrast, the athero-protective laminar flow suppressed YAP/TAZ activities. En face analysis of mouse arteries demonstrated an increased nuclear localization of YAP/TAZ and elevated levels of the target genes in the endothelium in atheroprone areas compared with athero-protective areas. YAP/TAZ knockdown significantly attenuated the disturbed flow induction of EC proliferative and proinflammatory phenotypes, whereas overexpression of constitutively active YAP was sufficient to promote EC proliferation and inflammation. In addition, treatment with statin, an antiatherosclerotic drug, inhibited YAP/TAZ activities to diminish the disturbed flow-induced proliferation and inflammation. In vivo blockade of YAP/TAZ translation by morpholino oligos significantly reduced endothelial inflammation and the size of atherosclerotic lesions. Our results demonstrate a critical role of the activation of YAP/TAZ by disturbed flow in promoting atheroprone phenotypes and atherosclerotic lesion development. Therefore, inhibition of YAP/TAZ activation is a promising athero-protective therapeutic strategy.

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Percentage Area of Intimal Surface of the Infra-Renal Segment of Abdominal Aorta Affected by Atherosclerosis - A Postmortem Study

Bangladesh Journal of Anatomy ◽

10.3329/bja.v7i1.3013 ◽

1970 ◽

Vol 7 (1) ◽

pp. 22-25

Author(s):

Saifun Nahar ◽

Humaira Naushaba ◽

Jesmin Ara Begum ◽

Naznin Akhter

Keyword(s):

Abdominal Aorta ◽

Age Groups ◽

Atherosclerotic Lesion ◽

Cross Sectional ◽

Medical College ◽

Atherosclerotic Lesions ◽

Significant Difference ◽

Percentage Area ◽

Present Rate ◽

Unpaired Student

Context: Atherosclerosis and its complications are still the major source of morbidity and mortality in the industrial world, and estimates have been offered that at its present rate of growth, will be the major cause of death from this disease by the year 2020 in the entire world, (Joseph 2004).The seemingly increased incidence of atherosclerotic disorders in the country, their fatal consequences and the paucity of relevant data, the present study was carried out to assess the percentage of area involved in atherosclerotic lesion of abdominal aorta of Bangladeshi males. Study design: Descriptive cross-sectional. Place & time of study: In the department of Anatomy, Sir Salimullah Medical College, Dhaka, during the study period of July 2005 to June 2006. Method: Abdominal aorta were collected from fifty Bangladeshi male dead bodies who underwent routine postmortem examination due to accidental death. Sudy was done to find out the macroscopic extent of atherosclerotic lesion of abdominal aorta (below the origin of renal arteries) from Bangladeshi male of different ages. The subjects were divided into lower (8-29), middle (30-40), higher (42-92) age groups. The atherosclerotic lesions were stained red by treating the aorta with sudden IV. The percentage of area on the intimal surface showing atherosclerotic lesion were measured by AutoCAD. Result: The percentage of area involved in atherosclerotic lesion was increase with advancing age. Unpaired student ‘t' test for statistical analysis revealed significant difference. (P<0.01) in between lower & higher age groups. Key words: Abdominal aorta, Atherosclerosis doi: 10.3329/bja.v7i1.3013 Bangladesh Journal of Anatomy January 2009, Vol. 7 No. 1 pp. 22-25

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Cellular composition of human early and advanced coronary atherosclerotic lesions

10.1101/2021.10.07.21264647 ◽

2021 ◽

Author(s):

Diana Sharysh ◽

Anton V. Markov ◽

Evgeniya S. Grigoryeva ◽

Olga E. Savelieva ◽

Aleksei A. Sleptcov ◽

...

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Flow Cytometry ◽

Atherosclerotic Lesion ◽

Interesting Case ◽

Atherosclerotic Plaques ◽

Cellular Composition ◽

Atherosclerotic Lesions ◽

A Cell ◽

First Time

AbstractObjectiveHere, we identify and quantify leukocytes, macrophages, endothelial cells (ECs), and vascular smooth muscle cells (VSMCs) with contractile and macrophage-like phenotypes by flow cytometry to compare human early and advanced coronary atherosclerotic plaques.Approach and ResultsSixteen coronary atherosclerotic lesions of 6 patients (3 women, 3 men, age 82 ± 9 years), including one case of restenosis after coronary stenting, were collected at autopsy. The cause of death of all patients was acute myocardial infarction. The lesions were categorized into early (EALs, n = 5) and advanced (AALs, n = 11) atherosclerotic stages. We analyzed a cell suspension stained with antibodies against CD45, CD68, CD31, and αSMA (ACTA2). We noted a decrease in the number of CD45+ cells and an increase in the CD45+CD68+ subpopulation of leukocytes from EALs to AALs. Numbers of CD45−αSMA+CD68+ cells positively correlated with the CD45+CD68+ macrophage number (r = 0.81; ρ = 0.64; p < 0.05) and the histological type of an atherosclerotic lesion (r = 0.81; ρ = 0.87; p < 0.05). As an interesting case, we analyzed cellular composition of the stented coronary artery and revealed significantly greater numbers of macrophages, αSMA+CD68+ VSMCs, and ECs in comparison with nonstented plaques.ConclusionsHuman early and advanced coronary atherosclerotic lesions differ in their counts of leukocytes and leukocyte subpopulations. For the first time, αSMA+CD68+ VSMCs were identified in early atherosclerotic stages of coronary arteries. Additionally, the restenotic coronary lesion contains mostly cells and is enriched in ECs, macrophages, and αSMA+CD68+ VSMCs in particular.

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Abstract 593: Hypoxia is Present in the Macrophage-rich Center of Human Carotid Atherosclerotic Plaques

Circulation ◽

10.1161/circ.116.suppl_16.ii_107-d ◽

2007 ◽

Vol 116 (suppl_16) ◽

Author(s):

Judith C Sluimer ◽

Job L van Wanroij ◽

Matthijs Groeneweg ◽

Bradly G Wouters ◽

Mat J Daemen ◽

...

Keyword(s):

Hypoxic Exposure ◽

Atherosclerotic Plaques ◽

List Type ◽

Strongly Correlated ◽

Plaque Instability ◽

Atherosclerotic Lesions ◽

Hypoxia Marker ◽

Diffuse Intimal Thickening ◽

Human Atherosclerosis ◽

The Media

Intraplaque neovascularization is linked to plaque instability and thought to be stimulated by hypoxia. However, hypoxia has not been demonstrated yet in human atherosclerosis. The hypoxia marker pimonidazole was administrated intravenously 2 hours prior to carotid endarterectomy in 6 symptomatic patients to evaluate the presence of hypoxia. Subsequent immunohistochemistry of the operatively removed atherosclerotic plaques demonstrated the presence of hypoxia, especially in the macrophage-rich center of the lesions. Notably, two hypoxic gradients were observed: hypoxia was very strong in the center of the plaque, but almost absent close to the main artery lumen and in the media. hypoxia was most intense in segments with advanced atheroma and almost absent in segments containing only diffuse intimal thickening. Hypoxia strongly correlated with CD68 immunoreactivity (ρ= 0.7, p=0.000), neovascularization (ρ= 0.6, p=0.000) and the presence of a thrombus (ρ= 0.4, p=0.009). In addition, hypoxia co-localized with expression of HIF1α and VEGF . To exclude that pimonidazole immunoreactivity in the atherosclerotic plaque was the result of surgery-induced ischemia, arterial wall segments were collected at two time-points: directly after incision of the carotid artery and directly following excision of the plaque. Pimonidazole immunoreactivity in these two pieces was not different, suggesting that hypoxia and pimonidazole adducts were already present in the plaques before surgery . To show that pimonidazole reactivity was hypoxia-specific and independent of reactive oxygen species, human THP-1 macrophages were exposed to normoxia (20% O 2 ), hypoxia (0.2% O 2 ) and/or H 2 O 2 (100 μM) in the presence of pimonidazole. Indeed, flow cytometry only showed pimonidazole-positive cells after hypoxic exposure. This is the first study proving direct evidence of the existence of hypoxia in advanced human atherosclerotic lesions, most prominently in the macrophage-rich center. Also, hypoxia was associated with the expression of HIF1α, VEGF and intraplaque microvessels, suggesting its involvement in the regulation of human intraplaque neovascularization.

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Reduction of oxidative stress and atherosclerosis in hyperlipidemic rabbits by Dioscorea rhizome

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y05-028 ◽

2005 ◽

Vol 83 (5) ◽

pp. 423-430 ◽

Cited By ~ 18

Author(s):

Weng-Cheng Chang ◽

Ya-Mei Yu ◽

Chieh-Hsi Wu ◽

Yueh-He Tseng ◽

Kuen-Yuh Wu

Keyword(s):

Oxidative Stress ◽

Antioxidant Enzymes ◽

Corn Oil ◽

Atherosclerotic Lesion ◽

Plasma Lipid ◽

Control Group ◽

Atherosclerotic Lesions ◽

Luminol Chemiluminescence ◽

Powdered Form ◽

Lucigenin Chemiluminescence

Hyperlipidemia may induce oxidative stress, which is important in the pathogenesis of atherosclerosis. Dioscorea rhizome (DR) is the powdered form of yams, and possesses antioxidant and hypolipidemic function. We therefore investigated the antioxidative and antiatherogenic effects of DR on hyperlipidemic rabbits. The control group was fed chow containing 0.5% cholesterol and 10% corn oil. The probucol and DR groups were fed the same diet as the control group but with the addition of 100 mg probucol/kg chow and 200 mg DR/kg chow, respectively. Total cholesterol and triacylglycerol plasma levels, RBC hemolysis T50, lucigenin chemiluminescence, and luminol chemiluminescence increased in the control group compared with the normal group, and decreased in the probucol and DR groups compared with the control group. The activity of antioxidant enzymes superoxide dismutase and catalase was significantly higher in the probucol and DR group than in the control group. The level of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in liver DNA was lower in the probucol and DR group than in the control group. Eighty percent of the intimal surface of the thoracic aorta was covered with atherosclerotic lesions in the control group but only 40% of the surface was covered in the DR group. These results suggest that supplementation with DR reduces oxidative stress and attenuates atherosclerosis in hyperlipidemic rabbits.Key words: Dioscorea rhizome, plasma lipid, RBC hemolysis, chemiluminescence, antioxidant enzymes, 8-OhdG, atherosclerotic lesion.

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En face dual fluorescence staining of fatty streaks allows observation of initiation and progression of atherosclerotic lesions

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100140695 ◽

1995 ◽

Vol 53 ◽

pp. 864-865

Author(s):

Anne M. Klinkner ◽

Crystal R. Waites ◽

Peter J. Bugelski ◽

William D. Kerns

Keyword(s):

Fluorescent Dyes ◽

Nile Red ◽

Dimethyl Formamide ◽

High Cholesterol Diet ◽

Methods Development ◽

Atherosclerotic Lesions ◽

En Face ◽

Biochemical Evaluation ◽

Stock Solutions ◽

Dual Staining

A primary effort in the understanding of the progression of atherosclerotic disease has been methods development for visualization of the atherosclerotic plaque. We introduce a new method for the qualitative analysis of lipids in atherosclerotic fatty streaks which also retains those lipids for biochemical evaluation. An original aspect of the process is the ability to view an entire fatty streak en face, selectively stained for specific lipid classes within the lesion.New Zealand white rabbits were fed a high cholesterol diet(0.15%-0.3% for 14 wks). The aorta was removed and fixed in Carson's phosphate buffered formaldehyde followed by dual staining in the fluorescent dyes Nile red and filipin. Stock solutions of nile red(0.5mg/ml acetone) and filipin(2.5mg/ml dimethyl formamide) were prepared and kept at -20°C; all subsequent steps were at RT. 0.5cm × 1.0cm pieces of aorta were trimmed and adventitia removed. The pieces were then washed 3×15 min in PBS w/o CaMg, soaked in Nile red(NR)/filipin(Fl) stain(100(il NR stock + 200μl Fl stock in 10 ml PBS for 30 min, washed in PBS 3×30 min, rinsed with distilled water, mounted(Crystal Mount, Biomedia) and coverslipped and viewed by fluorescence microscopy.

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Knowledge of non-healthcare individuals towards cardiopulmonary resuscitation: a cross-sectional study in Riyadh City, Saudi Arabia

International Journal of Emergency Medicine ◽

10.1186/s12245-021-00335-y ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Reema M. Alhussein ◽

Mansoor M. Albarrak ◽

Abdulaziz A. Alrabiah ◽

Nawfal A. Aljerian ◽

Hashim M. Bin Salleeh ◽

...

Keyword(s):

Health Care ◽

Saudi Arabia ◽

Cardiopulmonary Resuscitation ◽

Cross Sectional Study ◽

Sectional Study ◽

Cross Sectional ◽

Major Barrier ◽

Riyadh City ◽

Resuscitation Training ◽

The Media

Abstract Background Most sudden cardiac arrests occur at home, with low rates of bystander cardiopulmonary resuscitation being performed. We aimed to assess knowledge of cardiopulmonary resuscitation among individuals in Riyadh City, Saudi Arabia, who are not involved in health care. Methods A community-based cross-sectional study was conducted between January and February 2020 in 4 different areas in Riyadh City: North, South, East, and West. The participants were surveyed using a validated self-administered questionnaire. The Statistical Package for Social Sciences version 25.0 was used for inferential statistics and binary logistic regression analysis. Results A total of 856 participants completed the questionnaire, 51.8% were unaware of cardiopulmonary resuscitation. Only 4.4% of the participants had attended a formal cardiopulmonary resuscitation training course, 5.1% were campaign attendees, and 38.7% acquired their experience through the media. Having a higher level of education was positively associated with having knowledge of cardiopulmonary resuscitation. The main concern among attendees of cardiopulmonary resuscitation training courses and campaigns was legal issues, whereas inadequate knowledge was the major barrier for those who had learned about cardiopulmonary resuscitation through the media. Conclusion The level of knowledge of cardiopulmonary resuscitation among non-health care individuals in Riyadh City was found to be insufficient. Therefore, coordinated efforts among different authorities should be considered to implement a structured strategy aiming to increase awareness and knowledge of cardiopulmonary resuscitation among non-health care individuals.

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Comparison of swept-source versus spectral-domain optical coherence tomography angiography for detection of macular neovascularization

Graefe s Archive for Clinical and Experimental Ophthalmology ◽

10.1007/s00417-021-05229-6 ◽

2021 ◽

Author(s):

Anna Lentzsch ◽

Laura Schöllhorn ◽

Christel Schnorr ◽

Robert Siggel ◽

Sandra Liakopoulos

Keyword(s):

Optical Coherence Tomography ◽

Optical Coherence Tomography Angiography ◽

Spectral Domain ◽

Optical Coherence ◽

Cross Sectional ◽

Swept Source ◽

En Face ◽

Increased Sensitivity ◽

Sensitivity Specificity ◽

Subretinal Hyperreflective Material

Abstract Purpose To compare swept-source (SS) versus spectral-domain (SD) optical coherence tomography angiography (OCTA) for the detection of macular neovascularization (MNV). Methods In this prospective cohort study, 72 eyes of 54 patients with subretinal hyperreflective material (SHRM) and/or pigment epithelial detachment (PED) on OCT possibly corresponding to MNV in at least one eye were included. OCTA scans were acquired using two devices, the PLEX Elite 9000 SS-OCTA and the Spectralis SD-OCTA. Fluorescein angiography (FA) was used as reference. Two graders independently evaluated en face OCTA images using a preset slab as well as a manually modified slab, followed by a combination of en face and cross-sectional OCTA. Results Sensitivity (specificity) for the automated slabs was 51.7% (93.0%) for SS-OCTA versus 58.6% (95.3%) for SD-OCTA. Manual modification of segmentation increased sensitivity to 79.3% for SS-OCTA but not for SD-OCTA (58.6%). The combination of en face OCTA with cross-sectional OCTA reached highest sensitivity values (SS-OCTA: 82.8%, SD-OCTA: 86.2%), and lowest number of cases with discrepancies between SS-OCTA and SD-OCTA (4.2%). Fleiss kappa as measure of concordance between FA, SS-OCTA, and SD-OCTA was 0.56 for the automated slabs, 0.60 for the manual slabs, and 0.73 (good agreement) for the combination of en face OCTA with cross-sectional OCTA. Concordance to FA was moderate for the automated slabs and good for manual slabs and combination with cross-sectional OCTA of both devices. Conclusion Both devices reached comparable results regarding the detection of MNV on OCTA. Sensitivity for MNV detection and agreement between devices was best when evaluating a combination of en face and cross-sectional OCTA.

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Comparison of Osmotic Resistance, Shape and Transmembrane Potential of Erythrocytes Collected from Healthy and Fed with High Fat-Carbohydrates Diet (HF-CD) Pigs—Protective Effect of Cistus incanus L. Extracts

Materials ◽

10.3390/ma14041050 ◽

2021 ◽

Vol 14 (4) ◽

pp. 1050

Author(s):

Sylwia Cyboran-Mikołajczyk ◽

Robert Pasławski ◽

Urszula Pasławska ◽

Kacper Nowak ◽

Michał Płóciennik ◽

...

Keyword(s):

Biological Activity ◽

Physical Properties ◽

Plasma Lipid ◽

Membrane Lipid ◽

Osmotic Resistance ◽

High Fat ◽

Atherosclerotic Lesions ◽

Cistus Incanus ◽

Osmotic Resistance Of Erythrocytes ◽

The Impact

Long-term high fat-carbohydrates diet (HF-CD) contributes to the formation of irreversible changes in the organism that lead to the emergence of civilization diseases. In this study, the impact of three-month high-fat diet on the physical properties of erythrocytes (RBCs) was studied. Furthermore, the biological activity of Cistus incanus L. extracts, plant known with high pro-health potential, in relation to normal and HF-CD RBCs, was determined. Obtained results have shown that, applied HF-CD modified shape, membrane potential and osmotic resistance of erythrocytes causing changes in membrane lipid composition and the distribution of lipids. The impact of HF-CD on physical properties of RBCs along with atherosclerotic lesions of the artery was visible, despite the lack of statistically significant changes in blood morphology and plasma lipid profile. This suggests that erythrocytes may be good markers of obesity-related diseases. The studies of biological activity of Cistus incanus L. extracts have demonstrated that they may ameliorate the effect of HF-CD on erythrocytes through the membrane-modifying and antioxidant activity.

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