Cyclic fluctuations of 3‐hydroxy‐3‐methylglutaryl‐CoA reductase in aortic smooth muscle cell cultures

Alejandre, M.J.; Perales, Sonia; Carazo, Ángel; Palomino-Morales, Rogelio; Linares, Ana María

doi:10.1007/s11745-006-5058-x

Cited by 4 publications

(3 citation statements)

References 59 publications

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“…We have studied the cholesterol synthesis and HMG-CoA reductase gene expression in these cultures, showing great differences between SMC-C and SMC-Ch [50, 51]. Also we demonstrated the existence of cyclic fluctuations of HMG-CoA reductase activity in nonsynchronized SMC cultures not correlated to the cultured feeding and not with the increase of mRNA, suggesting the posttranscriptional modulation of the HMG-CoA reductase and the relationships between HMG-CoA reductase activity and cell division [52]. Finally, we have examined the morphological, molecular, and proliferation change in arterial SMC mimicking such a cholesterol diet.…”

Section: Discussionmentioning

confidence: 85%

Effect of Oxysterol‐Induced Apoptosis of Vascular Smooth Muscle Cells on Experimental Hypercholesterolemia

Perales

Alejandre

Palomino-Morales

et al. 2009

BioMed Research International

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Smooth muscle cells (SMCs) undergo changes related to proliferation and apoptosis in the physiological remodeling of vessels and in diseases such as atherosclerosis and restenosis. Recent studies also have demonstrated the vascular cell proliferation and programmed cell death contribute to changes in vascular architecture in normal development and in disease. The present study was designed to investigate the apoptotic pathways induced by 25-hydroxycholesterol in SMCs cultures, using an in vivo/in vitro cell model in which SMCs were isolated and culture from chicken exposed to an atherogenic cholesterol-rich diet (SMC-Ch) and/or an antiatherogenic fish oil-rich diet (SMC-Ch-FO). Cells were exposed in vitro to 25-hydroxycholesterol to study levels of apoptosis and apoptotic proteins Bcl-2, Bcl-XL and Bax and the expression of bcl-2 and bcl-xL, genes. The quantitative real-time reverse transcriptase-polymerase chain reaction and the Immunoblotting western blot analysis showed that 25-hydroxycholesterol produces apoptosis in SMCs, mediated by a high increase in Bax protein and Bax gene expression. These changes were more marked in SMC-Ch than in SMC-Ch-FO, indicating that dietary cholesterol produces changes in SMCs that make them more susceptible to 25-hydroxycholesterol-mediated apoptosis. Our results suggest that the replacement of a cholesterol-rich diet with a fish oil-rich diet produces some reversal of cholesterol-induced changes in the apoptotic pathways induced by 25-hydroxycholesterol in SMCs cultures, making SMCs more resistant to apoptosis.

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Section: Discussionmentioning

confidence: 85%

Effect of Oxysterol‐Induced Apoptosis of Vascular Smooth Muscle Cells on Experimental Hypercholesterolemia

Perales

Alejandre

Palomino-Morales

et al. 2009

BioMed Research International

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“…It has proven suitable for the in vitro study of the transformation into foam cells of SMCs that is induced in vivo by cholesterol diet, isolating the cells before plaque formation . Previously published experiments from our laboratory using this in vivo/in vitro model implicated cholesterol intake in the production of atherosclerotic lesions are consistent with the SMC modulation from contractile to synthetic state. In this study, we investigated the effect of cholesterol loading in vivo and in vitro on the gene expression of various ECM molecules.…”

Section: Discussionmentioning

confidence: 83%

Cholesterol loading in vivo and in vitro alters extracellular matrix proteins production in smooth muscle cells

Palomino-Morales

Perales

Torres

et al. 2015

Euro J Lipid Sci & Tech

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This study supports the SMC dedifferentiation in vitro model and increases our fundamental knowledge of plaque formation and remodeling. With this purpose, we isolated smooth muscle cells form aortic arch of chicks either fed under control or high cholesterol diets. From the former we obtained control SMCs (SMC−C) that were then loaded with cholesterol using cholesterol:methyl‐β‐cyclodextrin (CS) complexes, thus producing cholesterol loaded SMCs in vitro (SMC−C+CS).tained in vivo cholesterol loaded SMC cell cultures (SMC−Ch). We compared the effect of both cholesterol loading processes on extracellular matrix (ECM) production. The proline incorporation showed that collagen synthesis was higher in SMC−Ch and SMC−C+CS than in SMC−C cultures. Quantitative real‐time PCR revealed that expression of extracellular matrix protein‐related genes increased after in vitro and in vivo cholesterol loading with respect to unloaded cells, demonstrating a regulation at mRNA level. In addition, the expression of col3a1 and fibronectin genes was significantly higher in SMC−Ch than in SMC−Ch+CS cultures. We demonstrated that SMC can switch from contractile to synthetic state not only by cholesterol in vivo but also by cholesterol SMC loading in vitro. Practical applications: Alteration of the differentiated, contractile phenotype of SMCs is known to play an integral role in the formation of atherosclerotic lesions. Although SMCs have previously been induced to accumulate cholesterol in vitro, for either in vitro or in vivo foam cell like SMC cells, there is little information about phenotypic changes at the protein or RNA level. We have characterized the phenotypic changes associated with chicken aortic cholesterol loaded SMCs formation at the protein and molecular level. The results of the present study support the SMC dedifferentiation in vitro model and increase our fundamental knowledge of plaque formation and remodeling. Diet cholesterol is a risk factor in the atherosclerotic process. Proliferation, apoptosis, migration and the ECM synthesis in SMC contribute to the development of the disease. Both, a cholesterol‐rich diet and exogenous cholesterol loading in vitro yield the overexpression of ECM genes, specifically collagen types I and II and fibronectin. This leads to the dedifferentiation of the SMC to a synthetic phenotype which is one of the main reasons of the vascular hypertrophy.

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“…Hence, a high-cholesterol diet induced changes in the HMG-CoA reductase gene expression in aortic SMC of the chicks. The HMG-CoA reductase mRNA concentration in SMC cultures showed a marked rise after feeding that was not correlated with the fluctuation of activity observed during feeding of the cells for 72 h [34]. …”

Section: Introductionmentioning

confidence: 99%

Fish oil supplementation reverses the effect of cholesterol on apoptotic gene expression in smooth muscle cells

et al. 2010

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BackgroundNutritional control of gene regulation guides the transformation of smooth muscle cells (SMC) into foam cells in atherosclerosis. Oxidative stress has been reported in areas of lipid accumulation, activating proliferation genes. Suppression of oxidative stress by antioxidant administration reduces this activation and the progression of lesions. We hypothesized that fish oil consumption may protect against atherosclerotic vascular disease. The study objective was to determine the effects of dietary cholesterol and fish-oil intake on the apoptotic pathways induced by 25-hydroxycholesterol (25-HC) in SMC cultures.MethodsAn in vivo/in vitro cell model was used, culturing SMC isolated from chicks exposed to an atherogenic cholesterol-rich diet with 5% of cholesterol (SMC-Ch) alone or followed by an anti-atherogenic fish oil-rich diet with 10% of menhaden oil (SMC-Ch-FO) and from chicks on standard diet (SMC-C). Cells were exposed to 25-HC, studying apoptosis levels by flow cytometry (Annexin V) and expressions of caspase-3, c-myc, and p53 genes by quantitative real-time reverse transcriptase-polymerase chain reaction. Results: Exposure to 25-HC produced apoptosis in all three SMC cultures, which was mediated by increases in caspase-3, c-myc, and p53 gene expression. Changes were more marked in SMC-Ch than in SMC-C, indicating that dietary cholesterol makes SMC more susceptible to 25-HC-mediated apoptosis. Expression of p53 gene was elevated in SMC-Ch-FO. This supports the proposition that endogenous levels of p53 protect SMC against apoptosis and possibly against the development of atherosclerosis. Fish oil attenuated the increase in c-myc levels observed in SMC-C and SMC-Ch, possibly through its influence on the expression of antioxidant genes.ConclusionReplacement of a cholesterol-rich diet with a fish oil-rich diet produces some reversal of the cholesterol-induced changes, increasing the resistance of SMC to apoptosis.

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Cyclic fluctuations of 3‐hydroxy‐3‐methylglutaryl‐CoA reductase in aortic smooth muscle cell cultures

Cited by 4 publications

References 59 publications

Effect of Oxysterol‐Induced Apoptosis of Vascular Smooth Muscle Cells on Experimental Hypercholesterolemia

Effect of Oxysterol‐Induced Apoptosis of Vascular Smooth Muscle Cells on Experimental Hypercholesterolemia

Cholesterol loading in vivo and in vitro alters extracellular matrix proteins production in smooth muscle cells

Fish oil supplementation reverses the effect of cholesterol on apoptotic gene expression in smooth muscle cells

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