Changes in cultured arterial smooth muscle cells isolated from chicks upon cholesterol feeding

Carazo, Ángel; Alejandre, M.J.; Díaz, Ramón; Rı́os, Antonio; Castillo, M.; Linares, Ana María

doi:10.1007/s11745-998-0194-x

Cited by 17 publications

(26 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Twenty day-old chicken embryo pulmonary arterial smooth muscle cells (PASMCs) were isolated by enzymatic digestion as previously described [19,20]. In brief, proximal pulmonary arteries were isolated under aseptic condition and were cut into small pieces after removal of fat, adventitia, and connective tissue surrounding the arteries.…”

Section: Methodsmentioning

confidence: 99%

Hypoxia promotes cell proliferation by modulating E2F1 in chicken pulmonary arterial smooth muscle cells

Yang

Sun

Zhang

et al. 2013

J Animal Sci Biotechnol

View full text Add to dashboard Cite

In this study, we sought to investigate the expression of the transcription factor E2F1 in chicken pulmonary arterial smooth muscle cells upon hypoxia exposure, as well as the role that E2F1 played in the regulation of cell proliferation. Isolated chicken pulmonary arterial smooth muscle cells were subjected to hypoxia or normoxia for indicated time points. Cell viability, DNA synthesis, cell cycle profile, and expression of E2F1 were analyzed. The results showed that hypoxia promoted cell proliferation and DNA synthesis which was accompanied by an increased S phase entry and upregulation of E2F1 at mRNA and protein levels. Using siRNA technology, we demonstrated that gene inactivation of endogenous E2F1 abolished hypoxia-induced cell proliferation, DNA synthesis, and S phase entry compared with negative siRNA transfected cells. These results suggest that hypoxia-induced proliferation is mediated by inducing E2F1 in chicken pulmonary arterial smooth muscle cells.

show abstract

Section: Methodsmentioning

confidence: 99%

Hypoxia promotes cell proliferation by modulating E2F1 in chicken pulmonary arterial smooth muscle cells

Yang

Sun

Zhang

et al. 2013

J Animal Sci Biotechnol

View full text Add to dashboard Cite

show abstract

“…SMCs were isolated from the aortic arch of the chicks as described elsewhere [17] with slight modifications and cultured in Dulbecco's Modification of Eagle's Medium (DMEM) supplemented with D-glucose (4.5 g/L), L-glutamate (0.584 g/mL), antibiotic cocktail composed of penicillin (100 μ g/mL) and amphotericin (0.25 μ g/mL) (Sigma-Aldrich, Inc.), as well as 10% (v/v) fetal bovine serum (FBS). Medium was buffered with bicarbonate and cultures were kept at 37°C in humidified atmosphere of 95% air and 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

“…Thus, chickens have been reported to readily develop hypercholesterolemia with a cholesterol-rich diet, producing atherosclerotic lesions in some cases [14–16]. Preliminary studies demonstrated that a 20-day diet enriched with 5% cholesterol is sufficient to cause hypercholesterolemia and gives rise to lipid deposits in the main arteries that are similar to the fatty streaks observed in the first stages of atherosclerosis [17]. …”

Section: Introductionmentioning

confidence: 99%

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

Perales

Alejandre

Palomino-Morales

et al. 2009

BioMed Research International

Self Cite

View full text Add to dashboard Cite

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.

show abstract

“…SMC were isolated from the aortic arch of the chicks as described elsewhere with slight modifications [31] and cultured in Dulbecco's Modification of Eagle's Medium (DMEM) supplemented with D-glucose (4.5 g/l), L-glutamate (0.584 g/ml), antibiotic cocktail composed of penicillin (100 mg/ml) and amphotericin (0.25 mg/ml) (Sigma-Aldrich, Inc), and 10% (v/v) fetal bovine serum (FBS). Medium was buffered with bicarbonate, and cultures were kept at 37°C in humidified atmosphere (95% air, 5% CO 2 ).…”

Section: Methodsmentioning

confidence: 99%

“…It has proven suitable for in vitro study of the transformation into foam cells of SMC induced in vivo by cholesterol diet, isolating the cells before plaque formation [31]. Previous results with this model showed marked differences in proliferation and synthesis of DNA, RNA and protein between Ch-SMC and C-SMC, and these cell lines differed in lipid (especially cholesterol) synthesis and in the effects of inhibition on the first step of cholesterol biosynthesis [31,32]. In another study, Ch-SMC and C-SMC cultures showed altered HMG-CoA reductase activity and gene expression at transcriptional level [33].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2010

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Changes in cultured arterial smooth muscle cells isolated from chicks upon cholesterol feeding

Cited by 17 publications

References 48 publications

Hypoxia promotes cell proliferation by modulating E2F1 in chicken pulmonary arterial smooth muscle cells

Hypoxia promotes cell proliferation by modulating E2F1 in chicken pulmonary arterial smooth muscle cells

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

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

Contact Info

Product

Resources

About