A Volumetric Method for Quantifying Atherosclerosis in Mice by Using MicroCT: Comparison to En Face

Lloyd, D. J.; Helmering, Joan; Kaufman, Stephen A.; Turk, James R.; Silva, Matthew J.; Vasquez, Sergio X.; Weinstein, David M.; Johnston, B. G.; Hale, Clarence; Véniant, Murielle M.

doi:10.1371/journal.pone.0018800

Cited by 22 publications

(21 citation statements)

References 29 publications

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“…Mice were killed and aortas from heart to the iliac arteries were dissected out, soaked for 24 h in 4% paraformaldehyde for fixation, excised longitudinally and then stained with 0.5% of Sudan IV as described previously (Lloyd et al 2011). After staining, the aortas were laid onto the sponge block surface with the intimal surface up and pined down using Minutien (Fine Science Tools, Inc., San Francisco, CA, USA).…”

Section: En Face Analysismentioning

confidence: 99%

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TLR4 antagonist reduces early-stage atherosclerosis in diabetic apolipoprotein E-deficient mice

Lu¹,

Zhang²,

Li³

et al. 2012

Journal of Endocrinology

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Although it has been reported that deficiency of toll-like receptor 4 (TLR4) is associated with reduced atherosclerosis in atherosclerosis-prone mice and attenuated pro-inflammatory state in diabetic mice, it remains undetermined whether treatment with a TLR4 antagonist reduces atherosclerosis in nondiabetic or diabetic mice that have TLR4 expression. In this study, we determined the effect of Rhodobacter sphaeroides lipopolysaccharide (Rs-LPS), an established TLR4 antagonist, on early-stage atherosclerosis in nondiabetic and streptozotocin-induced diabetic apolipoprotein E-deficient (Apoe K/K ) mice. Analysis of atherosclerotic lesions of both en face aortas and cross sections of aortic roots showed that administration of Rs-LPS in 14-week-old diabetic Apoe K/K mice for 10 weeks significantly reduced atherosclerotic lesions. Although atherosclerotic lesions in nondiabetic Apoe K/K mice appeared to be decreased by Rs-LPS treatment, the difference was not statistically significant. Metabolic study showed that Rs-LPS significantly lowered serum levels of cholesterol and triglycerides in nondiabetic mice but not in diabetic mice. Furthermore, immunohistochemistry studies showed that Rs-LPS inhibited the expression of interleukin 6 and matrix metalloproteinase-9 and reduced the content of monocytes and macrophages in atherosclerotic plaques. Taken together, this study demonstrated for the first time that TLR4 antagonist inhibited vascular inflammation and atherogenesis in diabetic Apoe K/K mice and lowered serum cholesterol and triglyceride levels in nondiabetic Apoe K/K mice.

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Section: En Face Analysismentioning

confidence: 99%

“…The method to use the 'Similar' feature to select a particular color staining on a digitized immunohistochemical image has previously been described in detail (Lloyd et al 2011). Briefly, a standard was created by selecting an area of 0.5!0.5 cm from a tissue section that had desired brown color from immunostaining.…”

Section: Image Analysismentioning

confidence: 99%

TLR4 antagonist reduces early-stage atherosclerosis in diabetic apolipoprotein E-deficient mice

Lu¹,

Zhang²,

Li³

et al. 2012

Journal of Endocrinology

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show abstract

“…Mice were euthanized and aortas from heart to the iliac arteries were dissected out, soaked for 24 h in 4% paraformaldehyde for fixation, excised longitudinally, and then stained with 0.5% of Sudan IV as described previously 32 . After staining, the aortas were laid onto the sponge block surface with the intimal surface up and pined down using the Minutien (Fine Science Tools, Inc., San Francisco, CA).…”

Section: Methodsmentioning

confidence: 99%

Cooperative stimulation of atherogenesis by lipopolysaccharide and palmitic acid-rich high fat diet in low-density lipoprotein receptor-deficient mice

Brinson

et al. 2017

Atherosclerosis

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Background and aims Either lipopolysaccharide (LPS) or high-fat diet (HFD) enriched with saturated fatty acid (SFA) promotes atherosclerosis. In this study, we investigated the effect of LPS in combination with SFA-rich HFD on atherosclerosis and how LPS and SFA interact to stimulate inflammatory response in vascular endothelial cells. Methods Low-density lipoprotein receptor-deficient (LDLR−/−) mice were fed low-fat diet (LFD), HFD with low palmitic acid (PA) (LP-HFD), or HFD with high PA (HP-HFD) for 20 weeks. During the last 12 weeks, half mice received LPS and half received PBS. After treatment, metabolic parameters and aortic atherosclerosis were analyzed. To understand the underlying mechanisms, human aortic endothelial cells (HAECs) were treated with LPS and/or PA and proinflammatory molecule expression was quantified. Results Metabolic study showed that LPS had no significant effect on cholesterol, triglycerides, free fatty acids, but increased insulin and insulin resistance. Both LP-HFD and HP-HFD increased body weight and cholesterol while LP-HFD increased glucose and HP-HFD increased triglycerides, insulin, and insulin resistance. Analysis of aortic atherosclerosis showed that HP-HFD was more effective than LP-HFD in inducing atherosclerosis and LPS in combination with HP-HFD increased atherosclerosis in the thoracic aorta, a less common site for atherosclerosis, as compared with LPS or HP-HFD. To understand the mechanisms, results showed that LPS and PA synergistically upregulated adhesion molecules and proinflammatory cytokines in HAECs. Conclusions LPS and PA-rich HFD cooperatively increased atherogenesis in the thoracic aorta. The synergy between LPS and PA on proinflammatory molecules in HAECs may play an important role in atherogenesis.

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“…Mice were euthanized and aortas from heart to the iliac arteries were dissected out, soaked for 24 h in 4% paraformaldehyde for fixation, excised longitudinally, and then stained with 0.5% of Sudan IV as described previously (Lloyd, et al, 2011). After staining, the aortas were laid onto the sponge block surface with the intimal surface up and pined down using the Minutien (Fine Science Tools, Inc., San Francisco, CA).…”

Section: Methodsmentioning

confidence: 99%

TLR4 antagonist attenuates atherogenesis in LDL receptor-deficient mice with diet-induced type 2 diabetes

Zhang

et al. 2015

Immunobiology

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Although a large number of studies have well documented a key role of toll-like receptor (TLR)4 in atherosclerosis, it remains undetermined if TLR4 antagonist attenuates atherogenesis in mouse model for type 2 diabetes. In this study, we induced type 2 diabetes in low-density lipoprotein receptor-deficient (LDLR−/−) mice by high-fat diet (HFD). At 8 weeks old, 20 mice were fed HFD and 20 mice fed regular chow (RC) for 24 weeks. In the last 10 weeks, half HFD-fed mice and half RC-fed mice were treated with Rhodobacter sphaeroides lipopolysaccharide (Rs-LPS), an established TLR4 antagonist. After the treatment, atherosclerotic lesions in aortas were analyzed. Results showed that the HFD significantly increased bodyweight, glucose, lipids including total cholesterol, triglycerides and free fatty acids, and insulin resistance, indicating that the HFD induced type 2 diabetes in LDLR−/− mice. Results also showed that Rs-LPS had no effect on HFD-increased metabolic parameters in both nondiabetic and diabetic mice. Lipid staining of aortas and histological analysis of cross-sections of aortic roots showed that diabetes increased atherosclerotic lesions, but Rs-LPS attenuated atherogenesis in diabetic mice. Furthermore, immunohistochemical studies showed that Rs-LPS reduced infiltration of monocytes/macrophages and expression of interleukin (IL)-6 and matrix metalloproteinase-9 in atherosclerotic lesions of diabetic mice. Finally, the antagonistic effect of Rs-LPS on TLR4 was demonstrated by our in vitro studies showing that Rs-LPS inhibited IL-6 secretion from macrophages and endothelial cells stimulated by LPS or LPS plus saturated fatty acid palmitate. Taken together, our study demonstrated that TLR4 antagonist was capable of attenuating vascular inflammation and atherogenesis in mice with HFD-induced type 2 diabetes.

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A Volumetric Method for Quantifying Atherosclerosis in Mice by Using MicroCT: Comparison to En Face

Cited by 22 publications

References 29 publications

TLR4 antagonist reduces early-stage atherosclerosis in diabetic apolipoprotein E-deficient mice

TLR4 antagonist reduces early-stage atherosclerosis in diabetic apolipoprotein E-deficient mice

Cooperative stimulation of atherogenesis by lipopolysaccharide and palmitic acid-rich high fat diet in low-density lipoprotein receptor-deficient mice

TLR4 antagonist attenuates atherogenesis in LDL receptor-deficient mice with diet-induced type 2 diabetes

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