Oil Red O and Hematoxylin and Eosin Staining for Quantification of Atherosclerosis Burden in Mouse Aorta and Aortic Root

Andrés‐Manzano, María Jesús; Andrés, Vicente; Dorado, Beatriz

doi:10.1007/978-1-4939-2929-0_5

Cited by 92 publications

(50 citation statements)

References 2 publications

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“…These opened up aortic trees were then stained with Oil-Red-O and images were taken using stereo-microscope equipped with a camera, as described previously 34 . The percentage of total area stained by Oil-Red-O was determined using the NIH Image J software 39 using methodology as previously described 33 , 40 . Lesion morphology and atherosclerosis development in the carotid arteries was studied initially by gross imaging of the carotid arteries followed by performing Oil-Red-O staining on the cross-sections of the RCAs and LCAs.…”

Section: Methodsmentioning

confidence: 99%

Accelerated atherosclerosis development in C57Bl6 mice by overexpressing AAV-mediated PCSK9 and partial carotid ligation

Kumar

Kang

Rezvan

et al. 2017

Laboratory Investigation

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Studying the role of a particular gene in atherosclerosis typically requires a time-consuming and often difficult process of generating double-knockouts or transgenics on ApoE−/− or LDL receptor−/− background. Recently, it was reported that adeno-associated-virus-8 (AAV8) mediated overexpression of PCSK9 (AAV8-PCSK9) rapidly induced hyperlipidemia. However, using this method in C57BL6 wild-type (C57) mice, it took approximately 3 months to develop atherosclerosis. Our partial carotid ligation model is used to rapidly develop atherosclerosis by inducing disturbed flow in the left common carotid artery within 2 weeks in ApoE−/− or LDLR−/− mice. Here, we combined these two approaches to develop an accelerated model of atherosclerosis in C57 mice. C57 mice were injected with AAV9-PCSK9 or AAV9-Luciferase (control) and high-fat diet was initiated. A week later, partial ligation was performed. Compared to the control, AAV-PCSK9 led to elevated serum PCSK9, hypercholesterolemia, and rapid atherosclerosis development within 3 weeks as determined by gross plaque imaging, and staining with Oil-Red-O, Movat’s pentachrome and CD45 antibody. These plaque lesions were comparable to the atherosclerotic lesions that have been previously observed in ApoE−/− or LDLR−/− mice that were subjected to partial carotid ligation and high-fat diet. Next, we tested whether our method can be utilized to rapidly determine the role of a particular gene in atherosclerosis. Using eNOS−/− and NOX1−/y mice on C57 background, we found that the eNOS−/− mice developed more advanced lesions, while the NOX1−/y mice developed less atherosclerotic lesions as compared to the C57 controls. These results are consistent with the previous findings using double knockouts (eNOS−/−_ApoE−/− and NOX1−/y_ApoE−/−). AAV9-PCSK9 injection followed by partial carotid ligation is an effective and time-saving approach to rapidly induce atherosclerosis. This accelerated model is well-suited to quickly determine the role of gene(s) interest without generating double- or triple-knockouts.

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Section: Methodsmentioning

confidence: 99%

Accelerated atherosclerosis development in C57Bl6 mice by overexpressing AAV-mediated PCSK9 and partial carotid ligation

Kumar

Kang

Rezvan

et al. 2017

Laboratory Investigation

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“…The distal end of the aortic sinus was identified by the disappearance of the three aortic valve cusps as previously described [52]. For quantitative analysis of AS, every sixth section (spacing 50 μm apart) from each mouse was stained with Oil Red O and observed under 40× magnification (Olympus IX81, Tokyo, Japan).…”

Section: Methodsmentioning

confidence: 99%

Folic Acid Supplementation Delays Atherosclerotic Lesion Development by Modulating MCP1 and VEGF DNA Methylation Levels In Vivo and In Vitro

Cui

et al. 2017

IJMS

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The pathogenesis of atherosclerosis has been partly acknowledged to result from aberrant epigenetic mechanisms. Accordingly, low folate levels are considered to be a contributing factor to promoting vascular disease because of deregulation of DNA methylation. We hypothesized that increasing the levels of folic acid may act via an epigenetic gene silencing mechanism to ameliorate atherosclerosis. Here, we investigated the atheroprotective effects of folic acid and the resultant methylation status in high-fat diet-fed ApoE knockout mice and in oxidized low-density lipoprotein-treated human umbilical vein endothelial cells. We analyzed atherosclerotic lesion histology, folate concentration, homocysteine concentration, S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH), and DNA methyltransferase activity, as well as monocyte chemotactic protein-1 (MCP1) and vascular endothelial growth factor (VEGF) expression and promoter methylation. Folic acid reduced atherosclerotic lesion size in ApoE knockout mice. The underlying folic acid protective mechanism appears to operate through regulating the normal homocysteine state, upregulating the SAM: SAH ratio, elevating DNA methyltransferase activity and expression, altering MCP1 and VEGF promoter methylation, and inhibiting MCP1 and VEGF expression. We conclude that folic acid supplementation effectively prevented atherosclerosis by modifying DNA methylation through the methionine cycle, improving DNA methyltransferase activity and expression, and thus changing the expression of atherosclerosis-related genes.

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“…Aortas were washed twice with 78% methanol followed by phosphate buffer solution. En face images of aortic arches were visualized under a stereomicroscope (Olympus SZ, OLYMPUS, Tokyo, Japan) at ×20 magnification [28].…”

Section: Assessment On the Presence Of Atherosclerotic Plaque In Aortmentioning

confidence: 99%

Phenolic Compounds and the Anti-Atherogenic Effect of Bee Bread in High-Fat Diet-Induced Obese Rats

et al. 2019

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This study was undertaken to determine the phenolic compounds and the anti-atherogenic effect of bee bread in high-fat diet (HFD)-induced obese rats. The presence of phenolic compounds in bee bread was determined by liquid chromatography–mass spectrometry. Thirty-two male Sprague Dawley rats were divided into four groups, (n = 8/group); i.e., Normal (N), HFD (high-fat diet), HFD + BB (high-fat diet and 0.5 g/kg/day bee bread), and HFD + O (high-fat diet and 10 mg/kg/day orlistat) groups. After 6 weeks of the experiment, rats were sacrificed. Five phenolic compounds were identified in bee bread; namely, caffeic acid, ferulic acid, kaempferol, apigenin, and isorhamnetin. Bee bread significantly reduced Lee obesity index and levels of total cholesterol (TC), low-density lipoprotein (LDL), fatty acid synthase (FAS) activity, atherogenic index, oxidised-LDL (oxLDL), and malondialdehyde (MDA), and significantly increased aortic antioxidant activities, such as those of superoxide dismutase (SOD) and glutathione peroxidase (GPx). Adipocyte sizes were found to be smaller in the HFD + BB group compared to the N group, and en face aortas showed an absence of atherosclerotic plaque in rats supplemented with bee bread. These changes might suggest an anti-atherogenic effect of bee bread in HFD-induced obese rats via its antioxidant and hypocholesterolaemic properties.

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Oil Red O and Hematoxylin and Eosin Staining for Quantification of Atherosclerosis Burden in Mouse Aorta and Aortic Root

Cited by 92 publications

References 2 publications

Accelerated atherosclerosis development in C57Bl6 mice by overexpressing AAV-mediated PCSK9 and partial carotid ligation

Accelerated atherosclerosis development in C57Bl6 mice by overexpressing AAV-mediated PCSK9 and partial carotid ligation

Folic Acid Supplementation Delays Atherosclerotic Lesion Development by Modulating MCP1 and VEGF DNA Methylation Levels In Vivo and In Vitro

Phenolic Compounds and the Anti-Atherogenic Effect of Bee Bread in High-Fat Diet-Induced Obese Rats

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