human aortic smcs (hasmcs  (CHI Scientific Inc)

Journal: Journal of the American Heart Association

Article Title: Brain‐Derived Neurotrophic Factor Expression and Signaling in Different Perivascular Adipose Tissue Depots of Patients With Coronary Artery Disease

doi: 10.1161/jaha.120.018322

Figure Lengend Snippet: Figure 6. Morphological and functional response of human aortic smooth muscle cells (HASMCs) to brain-derived neurotrophic factor (BDNF) stimulation and importance of protein tyrosine phosphatase 1B (PTP1B). HASMCs were cultivated on gelatin-coated coverslips and treated with PTP1B inhibitor (50 µmol/L) or dimethyl sulfoxide (DMSO) for 1 hour before being stimulated with recombinant human BDNF (10 ng/mL) for an additional 24 hours. Cells were fixed and changes in the expression of smooth muscle α-actin (SMA; A, top row) or MYH10 (A, bottom row) or proliferating cell nuclear antigen (PCNA; B, top row; arrows point to positive cells) and actin fiber arrangement (F-actin; B, bottom row) examined using confocal fluorescence microscopy. Scale bars indicate 40 µm (A and B, bottom row) or 20 µm (B, top row). Cell proliferation was analyzed using the CellTiter 96 AQueous One Solution cell proliferation assay (C) and cell proliferation and migration using the scratch-wound assay (D and E). *P<0.05, **P<0.01, and ****P<0.0001 vs dimethyl sulfoxide (DMSO) and ##P<0.01 vs BDNF alone, as determined using 1-way ANOVA followed by multiple comparisons test.

Article Snippet: Human aortic SMCs (HASMCs) were purchased from PromoCell (C- 12532) and cultivated in Smooth Muscle Cell Growth Medium 2 (PromoCell, C- 22062), containing 5% fetal calf serum as well as insulin (5 μg/mL), epidermal growth factor (0.5 ng/mL), and basic fibroblast growth factor (2 ng/mL), in a humidified 5% CO2 atmosphere at 37°C.

Techniques: Functional Assay, Derivative Assay, Recombinant, Expressing, Fluorescence, Microscopy, Proliferation Assay, Migration, Scratch Wound Assay Assay

Journal: Journal of the American Heart Association

Article Title: Brain‐Derived Neurotrophic Factor Expression and Signaling in Different Perivascular Adipose Tissue Depots of Patients With Coronary Artery Disease

doi: 10.1161/jaha.120.018322

Figure Lengend Snippet: Figure 7. Hypothetical signaling of perivascular neurotrophins in vascular smooth muscle cells (SMCs) during atherosclerosis and the effects of protein tyrosine phosphatase 1B (PTP1B) or its inhibition. Chronically increased expression of brain-derived neurotrophic factor (BDNF) as well as other adipokines (eg, leptin), inflammatory mediators (eg, tumor necrosis factgor α [TNF-α]), and counterregulatory phosphatases (eg, PTP1B), as observed in perivascular adipose tissue surrounding the aortic root and coronary arteries of patients with advanced atherosclerosis, is associated with downregulation of the BDNF receptor on SMCs. Negative regulation of tropomyosin kinase (Trk) B signaling by PTP1B may contribute to atherosclerosis by altering the effects of BDNF on SMC proliferation, migration, differentiation, or collagen production. Inhibition of PTP1B upregulates the low-affinity neurotrophin receptor p75NTR and inhibits the effects of BDNF on SMCs.

Article Snippet: Human aortic SMCs (HASMCs) were purchased from PromoCell (C- 12532) and cultivated in Smooth Muscle Cell Growth Medium 2 (PromoCell, C- 22062), containing 5% fetal calf serum as well as insulin (5 μg/mL), epidermal growth factor (0.5 ng/mL), and basic fibroblast growth factor (2 ng/mL), in a humidified 5% CO2 atmosphere at 37°C.

Techniques: Inhibition, Expressing, Derivative Assay, Migration

Journal: Journal of Cellular and Molecular Medicine

Article Title: MicroRNA‐92a promotes vascular smooth muscle cell proliferation and migration through the ROCK/MLCK signalling pathway

doi: 10.1111/jcmm.14274

Figure Lengend Snippet: Expression of KLF4 in blood vessel walls of AS mice and effects of siRNA‐KLF4 (siKLF4) on vascular smooth muscle cells (VSMCs) function. A, Immunofluorescence staining for KLF4 (red) and α‐SMA (green) in AS mice and ML‐7 treatment mice. Nuclei were stained with DAPI (blue). Bars, 50 μm. B, The proliferation of siKLF4‐transfected Rat Primary Aortic SMCs were measured by CCK8 with 10 ng/mL PDGF‐BB stimulation. C and D, The migration of siKLF4‐transfected Rat Primary Aortic SMCs (C) and A7r5 (D) were measured by Boyden Chamber assay with 10 ng/mL PDGF‐BB stimulation. Migrated cells in each high‐power field (HPF, 400×) were quantitated and the results are shown on the right. E and F, The cells proliferation and migration of HASMCs were measured by CCK8 (E) and Boyden Chamber assay (F) respectively. Migrated cells in each high‐power field (HPF, 400×) were quantitated and the results are shown on the right. Data are presented as mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human aortic SMCs (HASMCs) were purchased from Chi Scientific Inc (Catalog No. 7‐1562, Jiangsu, China).

Techniques: Expressing, Immunofluorescence, Staining, Transfection, Migration, Boyden Chamber Assay