Glucagon-like peptide-1 inhibits vascular smooth muscle cell dedifferentiation through mitochondrial dynamics regulation

Torres, Gloria; Morales, Pablo E.; García-Miguel, Marina; Norambuena-Soto, Ignacio; Cartes-Saavedra, Benjamín; Vidal-Peña, Gonzalo; Moncada-Ruff, David; Sanhueza–Olivares, Fernanda; Martín, Alejandra San; Chiong, Mario

doi:10.1016/j.bcp.2016.01.013

Cited by 46 publications

(23 citation statements)

References 49 publications

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“…Cells were transferred into an incubation chamber (37°C, 5% CO 2 ), and images were acquired using LSM-800 time-lapse confocal microscopy with ×40 oil immersion objective lens. Z-stacks of thresholder images were volume reconstituted using the VolumeJ plug-in (31). We can track changes in mitochondrial size that ranged from 0.039 to 0.873 μm 2 .…”

Section: Methodsmentioning

confidence: 99%

Metformin Suppresses Diabetes-Accelerated Atherosclerosis via the Inhibition of Drp1-Mediated Mitochondrial Fission

et al. 2016

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Metformin is a widely used antidiabetic drug that exerts cardiovascular protective effects in patients with diabetes. How metformin protects against diabetes-related cardiovascular diseases remains poorly understood. Here, we show that metformin abated the progression of diabetes-accelerated atherosclerosis by inhibiting mitochondrial fission in endothelial cells. Metformin treatments markedly reduced mitochondrial fragmentation, mitigated mitochondrial-derived superoxide release, improved endothelial-dependent vasodilation, inhibited vascular inflammation, and suppressed atherosclerotic lesions in streptozotocin (STZ)-induced diabetic ApoE−/− mice. In high glucose–exposed endothelial cells, metformin treatment and adenoviral overexpression of constitutively active AMPK downregulated mitochondrial superoxide, lowered levels of dynamin-related protein (Drp1) and its translocation into mitochondria, and prevented mitochondrial fragmentation. In contrast, AMPK-α2 deficiency abolished the effects of metformin on Drp1 expression, oxidative stress, and atherosclerosis in diabetic ApoE−/−/AMPK-α2−/− mice, indicating that metformin exerts an antiatherosclerotic action in vivo via the AMPK-mediated blockage of Drp1-mediated mitochondrial fission. Consistently, mitochondrial division inhibitor 1, a potent and selective Drp1 inhibitor, reduced mitochondrial fragmentation, attenuated oxidative stress, ameliorated endothelial dysfunction, inhibited inflammation, and suppressed atherosclerosis in diabetic mice. These findings show that metformin attenuated the development of atherosclerosis by reducing Drp1-mediated mitochondrial fission in an AMPK-dependent manner. Suppression of mitochondrial fission may be a therapeutic approach for treating macrovascular complications in patients with diabetes.

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Section: Methodsmentioning

confidence: 99%

Metformin Suppresses Diabetes-Accelerated Atherosclerosis via the Inhibition of Drp1-Mediated Mitochondrial Fission

et al. 2016

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“…The A7r5 cell line is widely used in vitro to study the physiology and pathophysiology of VSMCs [ 13 , 14 ]. Some studies demonstrated this cell line as the “contractile” phenotype [ 17 , 18 ], while others showed the A7r5 cell line is “synthetic” phenotype VSMCs [ 19 ].…”

Section: Discussionmentioning

confidence: 99%

“…The A7r5 cell line has been widely used in vitro to study the physiology and pathophysiology of VSMCs [ 13 , 14 ]. However, it has lost some VSMCs selectivity and has many differences from primary cultured VSMCs.…”

Section: Methodsmentioning

confidence: 99%

Primary Culture of Rat Aortic Vascular Smooth Muscle Cells: A New Method

et al. 2017

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BackgroundDeveloping a simple and efficient method of obtaining primary cultured VSMCs is necessary for basic cardiovascular research.Material/MethodsThe procedure of our new method mainly includes 6 steps: isolation of the aortic artery, removal of the fat tissue around the artery, separation of the media, cutting the media into small tissue blocks, transferring the tissue blocks to cell culture plates, and incubation until the cells reach confluence. The cells were identified as VSMCs by morphology and immunofluorescence. Then, VSMCs obtained by this new tissue explants method, the traditional tissue explants method, the enzyme digestion method, and A7r5 cell line were divided into 4 groups. The purity of cells was test by multiple fluorescent staining. Western blotting was used to investigate the phenotype of VSMCs obtained by different methods.ResultsCells began to grow out at about 8 days and became relatively confluent within 16 days. Compared with VSMCs from the traditional tissue explants method and enzyme digestion method or A7r5 cell line, VSMCs obtained by our method showed higher purity and manifested a more “contractile” phenotype characteristic.ConclusionsWe have conquered the disadvantages in the previous primary culture methods and established a simple and reliable way to isolate and culture rat aortic VSMCs with high purity and stability.

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“…The smooth muscle A7r5 cell line was obtained from the thoracic aorta of BDIX rat ( Rattus novergicus) [ 9 ]. The A7r5 cells were purchased from ATCC (CRL-1444) and were expanded in DMEM supplemented with 2 mM glutamine and 10% FBS as described [ 10 , 11 ]. Prior to experiments, culture media was replaced with DMEM 2 mM glutamine and 2% FBS and cells were cultured for further 24 h. Total serum deprivation induced autophagy ( S1 Fig ).…”

Section: Methodsmentioning

confidence: 99%

“…Proliferation of VSMC was determined using MTT assay [ 12 ] and [ 3 H]-thymidine incorporation [ 11 ]. A7r5 migration was evaluated by the wound healing and transwell assays in the presence of bromodeoxiuridine to inhibit cell proliferation [ 11 ].…”

Section: Methodsmentioning

confidence: 99%

Autophagy mediates tumor necrosis factor-α-induced phenotype switching in vascular smooth muscle A7r5 cell line

et al. 2018

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Vascular smooth muscle cells (VSMC) dedifferentiation from a contractile to a synthetic phenotype contributes to atherosclerosis. Atherosclerotic tissue has a chronic inflammatory component with high levels of tumor necrosis factor-α (TNF-α). VSMC of atheromatous plaques have increased autophagy, a mechanism responsible for protein and intracellular organelle degradation. The aim of this study was to evaluate whether TNF-α induces phenotype switching of VSMCs and whether this effect depends on autophagy. Rat aortic Vascular smooth A7r5 cell line was used as a model to examine the phenotype switching and autophagy. These cells were stimulated with TNF-α 100 ng/mL. Autophagy was determined by measuring LC3-II and p62 protein levels. Autophagy was inhibited using chloroquine and siRNA Beclin1. Cell dedifferentiation was evaluated by measuring the expression of contractile proteins α-SMA and SM22, extracellular matrix protein osteopontin and type I collagen levels. Cell proliferation was measured by [3H]-thymidine incorporation and MTT assay, and migration was evaluated by wound healing and transwell assays. Expression of IL-1β, IL-6 and IL-10 was assessed by ELISA. TNF-α induced autophagy as determined by increased LC3-II (1.91±0.21, p<0.001) and decreased p62 (0.86±0.02, p<0.05) when compared to control. Additionally, TNF-α decreased α-SMA (0.74±0.12, p<0.05) and SM22 (0.54±0.01, p<0.01) protein levels. Consequently, TNF-α induced migration (1.25±0.05, p<0.05), proliferation (2.33±0.24, p<0.05), and the secretion of IL-6 (258±53, p<0.01), type I collagen (3.09±0.85, p<0.01) and osteopontin (2.32±0.46, p<0.01). Inhibition of autophagy prevented all the TNF-α-induced phenotypic changes. TNF-α induces phenotype switching in A7r5 cell line by a mechanism that required autophagy. Therefore, autophagy may be a potential therapeutic target for the treatment of atherosclerosis.

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Glucagon-like peptide-1 inhibits vascular smooth muscle cell dedifferentiation through mitochondrial dynamics regulation

Cited by 46 publications

References 49 publications

Metformin Suppresses Diabetes-Accelerated Atherosclerosis via the Inhibition of Drp1-Mediated Mitochondrial Fission

Metformin Suppresses Diabetes-Accelerated Atherosclerosis via the Inhibition of Drp1-Mediated Mitochondrial Fission

Primary Culture of Rat Aortic Vascular Smooth Muscle Cells: A New Method

Autophagy mediates tumor necrosis factor-α-induced phenotype switching in vascular smooth muscle A7r5 cell line

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