primary rat vascular smooth muscle cells (vsmcs)  (Dawley Inc)

Journal: PLoS ONE

Article Title: Possible Dual Role of Decorin in Abdominal Aortic Aneurysm

doi: 10.1371/journal.pone.0120689

Figure Lengend Snippet: ( A ) Cultured rat vascular smooth muscle cells (VSMCs) were pre-treated with or without 0.4, 4, or 40 μg/ml decorin, then stimulated with or without 100 ng/ml lipopolysaccharide (LPS) for 48 h. Protein levels of MMP-9 and MMP-2 in the conditioned media were determined by gelatin zymography. ( B ) Representative results are shown. ( C-D ) Cultured rat VSMCs were pre-treated with or without 40 μg/ml decorin, then stimulated with or without 100 ng/ml LPS for 48 h. Quantitative analyses for MMP-9 ( C ) and MMP-2 ( D ) are shown. Data are mean ± SD. ** p <0.01 compared to Control; ## p <0.01 compared to LPS.

Article Snippet: Rat aortic vascular smooth muscle cells (VSMCs) derived from the medial layer of healthy rat aorta were purchased from Cell Applications, Inc (San Diego, CA, USA).

Techniques: Cell Culture, Zymography, Control

Journal: Redox Biology

Article Title: Matrix metalloproteinase-2-induced epidermal growth factor receptor transactivation impairs redox balance in vascular smooth muscle cells and facilitates vascular contraction

doi: 10.1016/j.redox.2018.07.005

Figure Lengend Snippet: MMP-2-induced increase in ROS concentrations in vascular smooth muscle cells is prevented by MMP inhibitors. ( A) ROS concentrations in vascular smooth muscle cells (A7r5 cells) assessed with dihydroethidium (DHE; 10 µmol/l) by flow cytometry after incubation with MMP-2 (16 nmol/l) for 10 or 30 min. ( B) Effects of antioxidant agents including apocynin 100 µmol/l (Apo), diphenyl iodonium (DPI; a flavoprotein inhibitor) 10 µmol/l, or PEG-catalase 3000 U/ml (PG-Cat, which catalyzes the breakdown of H 2 O 2 ) in both vehicle and in MMP-2-treated cells. Panel C shows the effects of MMP inhibitors doxycycline 100 µmol/l (Doxy) or GM6001 1 µmol/l in both vehicle and in MMP-2-treated cells. Data are shown as the mean ± SEM (n = 4/group). * P < 0.05 vs. Vehicle. ** P < 0.05 vs. Vehicle and 10 min. # P < 0.05 vs. Vehicle in MMP-2-treated cells.

Article Snippet: The Rattus norvegicus vascular smooth muscle cell (VSMC) line A7r5 obtained from American Type Culture Collection (ATCC CRL-1444) (Rockville, MD, USA) was maintained at 37 °C under an atmosphere of 5% CO 2 in culture flasks with Dulbecco ́s modified Eagle ́s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% antibiotic-antimycotic solution (Life technologies, Cat# 15240112).

Techniques: Flow Cytometry, Incubation

Journal: Redox Biology

Article Title: Matrix metalloproteinase-2-induced epidermal growth factor receptor transactivation impairs redox balance in vascular smooth muscle cells and facilitates vascular contraction

doi: 10.1016/j.redox.2018.07.005

Figure Lengend Snippet: MMP-2 mediates HB-EGF shedding and induces EGFR-dependent increases in ROS concentrations . ( A) MMP-2 promotes HB-EGF shedding, as revealed by quantification of HB-EGF shedding measured by alkaline phosphatase (AP) activity in the supernatant of HEK293 cells after incubation of MMP-2 (16 nmol/l) or Vehicle for 10 and 30 min. Shedding activity is presented as the ratio of the AP activity in the supernatant and in the cell lysate as described in . ( B) Incubation with MMP-2 (16 nmol/l) for 10 or 30 min increases ROS concentrations in vascular smooth muscle cells (A7r5 cells) assessed with dihydroethidium (DHE; 10 µmol/l) by flow cytometry, and this effect is prevented by the EGFR kinase inhibitor (Ag1478; 3 or 10 μmol/l). Data are shown as the mean ± SEM (n = 4/group). * P < 0.05 vs . respective Control (Vehicle). * P < 0.05 vs. respective Control (Vehicle) and vs. MMP-2 10 min. # P < 0.05 vs. Vehicle + Vehicle treated cells. ## P < 0.05 vs. Vehicle + Vehicle treated cells and vs. MMP-2 + Vehicle treated cells for 10 min.

Article Snippet: The Rattus norvegicus vascular smooth muscle cell (VSMC) line A7r5 obtained from American Type Culture Collection (ATCC CRL-1444) (Rockville, MD, USA) was maintained at 37 °C under an atmosphere of 5% CO 2 in culture flasks with Dulbecco ́s modified Eagle ́s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% antibiotic-antimycotic solution (Life technologies, Cat# 15240112).

Techniques: Activity Assay, Incubation, Flow Cytometry, Control

Journal: Redox Biology

Article Title: Matrix metalloproteinase-2-induced epidermal growth factor receptor transactivation impairs redox balance in vascular smooth muscle cells and facilitates vascular contraction

doi: 10.1016/j.redox.2018.07.005

Figure Lengend Snippet: MMP-2-induced increases in ROS concentrations in vascular smooth muscle cells is abolished by a phospholipase C inhibitor or by a protein kinase C inhibitor . (A) Quantification of ROS production after 30 min of MMP-2 incubation with VSMC in the absence or presence of the phospholipase C inhibitor (A778 10 μM) or the PKC inhibitor (Chelerythrine, Chel, 3 μM). ROS concentrations were assessed with DHE probe by flow cytometry. (B) Control experiments showing quantification of ROS production stimulated by an EGFR agonist (recombinant HB-EGF) or by a PKC activator (phorbol 12-myristate 13-acetate; PMA) after 30 min of incubation with MMP-2. Data are shown as the mean ± SEM (n = 4/group). * P < 0.05 vs. Vehicle + Vehicle treated cells. # P < 0.05 vs. Vehicle + MMP-2 treated cells.

Article Snippet: The Rattus norvegicus vascular smooth muscle cell (VSMC) line A7r5 obtained from American Type Culture Collection (ATCC CRL-1444) (Rockville, MD, USA) was maintained at 37 °C under an atmosphere of 5% CO 2 in culture flasks with Dulbecco ́s modified Eagle ́s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% antibiotic-antimycotic solution (Life technologies, Cat# 15240112).

Techniques: Incubation, Flow Cytometry, Control, Recombinant