Differential effects of cadmium on proteoglycan synthesis of arterial smooth muscle cells: increase in small dermatan sulfate proteoglycans, biglycan and decorin, in the extracellular matrix at low cell density

Toxicology ◽  
2002 ◽  
Vol 170 (1-2) ◽  
pp. 89-101 ◽  
Author(s):  
Yasuyuki Fujiwara ◽  
Naoko Tsumura ◽  
Chika Yamamoto ◽  
Toshiyuki Kaji
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Cell Density ◽  
Dermatan Sulfate ◽  
Muscle Cells ◽  
Proteoglycan Synthesis ◽  
Differential Effects ◽  
Arterial Smooth Muscle Cells ◽  
Low Cell Density

scholarly journals Arsenite but not arsenate inhibits general proteoglycan synthesis in cultured arterial smooth muscle cells

2008 ◽  
Vol 33 (4) ◽  
pp. 487-492 ◽  
Author(s):  
Yasuyuki Fujiwara ◽  
Chika Yamamoto ◽  
Takashi Hirooka ◽  
Naoko Terada ◽  
Masahiko Satoh ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Proteoglycan Synthesis ◽  
Arterial Smooth Muscle ◽  
Arterial Smooth Muscle Cells

scholarly journals Inhibition of proteoglycan synthesis alters extracellular matrix deposition, proliferation, and cytoskeletal organization of rat aortic smooth muscle cells in culture.

1989 ◽  
Vol 108 (6) ◽  
pp. 2495-2505 ◽  
Author(s):  
H F Hamati ◽  
E L Britton ◽  
D J Carey
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Immunofluorescent Staining ◽  
Proteoglycan Synthesis ◽  
Aortic Smooth Muscle Cells ◽  
Aortic Smooth Muscle ◽  
Matrix Deposition ◽  
Alpha Actin

Arterial proteoglycans have been implicated in several important physiological processes ranging from lipid metabolism to regulation of smooth muscle cell growth. Vascular smooth muscle (VSM) cells are the major producers of proteoglycans in the medial layer of blood vessels. To study functional consequences of alterations in VSM proteoglycan metabolism we used 4-methylumbelliferyl-beta-D-xyloside to inhibit proteoglycan synthesis in primary and early passage cultures of rat aortic smooth muscle cells. Biochemical analysis of cultures labeled with 35SO4 showed the drug inhibited synthesis of different classes of proteoglycans by 50 to 62%. Inhibition of proteoglycan synthesis resulted in reduced accumulation of extracellular matrix, as shown by immunofluorescent staining with antibodies to chondroitin sulfate, fibronectin, thrombospondin, and laminin. There was also an inhibition of postconfluent (multilayered) growth of the smooth muscle cells, and a change in the morphology of the cells, with no apparent effect on subconfluent growth. In addition, in drug-treated cells there was a reduction in the number of cytoskeletal filaments that contained alpha-actin, the actin subtype synthesized by differentiated VSM cells. This occurred even though the total content of alpha-actin in the cells was not reduced. The effects of the inhibitor on growth and morphology could be reversed by switching the cultures to normal medium and could be prevented by growing the cells on preformed VSM extracellular matrix. These observations suggest the vascular extracellular matrix may play a role in regulating the growth and differentiation of smooth muscle cells.

scholarly journals Proteoglycans in primate arteries. III. Characterization of the proteoglycans synthesized by arterial smooth muscle cells in culture.

1983 ◽  
Vol 96 (1) ◽  
pp. 167-176 ◽  
Author(s):  
T N Wight ◽  
V C Hascall
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Chondroitin Sulfate ◽  
Dermatan Sulfate ◽  
Cell Layer ◽  
Muscle Cells ◽  
Macaca Nemestrina ◽  
Arterial Smooth Muscle ◽  
Dermatan Sulfate Proteoglycan ◽  
Arterial Smooth Muscle Cells

Near confluent monolayers of arterial smooth muscle cells derived from Macaca nemestrina were labeled with Na2[35S]O4 and the newly synthesized proteoglycans present in the culture medium and cell layer were extracted with either 4 M guanidine HCl (dissociative solvent) or 0.5 M guanidine HCl (associative solvent) in the presence of protease inhibitors. The proteoglycans in both compartments were further purified by cesium chloride density gradient ultracentrifugation. Two size classes of proteoglycans were observed in the medium as determined by chromatography on Sepharose CL-2B. The large population (Kav = 0.31) contained predominantly chondroitin sulfate chains with Mr = approximately 40,000. The smaller population (Kav = 0.61) contained dermatan sulfate chains of similar Mr (approximately 40,000). When tested for their ability to aggregate, only proteoglycans in the large-sized population were able to aggregate. A chondroitin sulfate containing proteoglycan with identical properties was isolated from the cell layer. In addition, the cell layer contained a dermatan sulfate component which eluted later on Sepharose CL-2B (Kav = 0.78) than the dermatan sulfate proteoglycan present in the medium. Electron microscopy of the purified proteoglycans revealed a bottlebrush structure containing a central core averaging 140 nm in length with an average of 8 to 10 side projections. The length of the side projections varied but averaged between 70 and 75 nm. Similar bottlebrush structures were observed in the intercellular matrix of the smooth muscle cell cultures after staining with Safranin 0. This culture system provides a model to investigate parameters involved in the regulation of synthesis and degradation of arterial proteoglycans.

scholarly journals Proteoglycans in primate arteries. II. Synthesis and secretion of glycosaminoglycans by arterial smooth muscle cells in culture.

1975 ◽  
Vol 67 (3) ◽  
pp. 675-686 ◽  
Author(s):  
T N Wight ◽  
R Ross
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Culture Medium ◽  
Dermatan Sulfate ◽  
Muscle Cells ◽  
Arterial Smooth Muscle ◽  
Glycosaminoglycan Synthesis ◽  
Arterial Smooth Muscle Cells ◽  
Chondroitin Sulfate A

Glycosaminoglycan synthesis and secretion by primate arterial smooth muscle have been examined in cell culture. Mass cultures of diploid primate arterial smooth muscle cells were either double labeled with [35S]sulfate and [3H]acetate or single labeled with [3H]glucosamine for 24 h and glycosaminoglycans were extracted and isolated from the culture medium. Incorporation of labeled precursors into glycosaminoglycan was maximal during stationary phase of smooth muscle cell growth in culture and reduced, but not eliminated during logarithmic growth. The glycosaminoglycans synthesized and secreted into the culture medium were characterized by differential susceptibility to glycosaminoglycan-degradative enzymes and by cellulose acetate electrophoresis. Both assay procedures indicate that cultured primate arterial smooth muscle cells synthesize principally dermatan sulfate (60%-80% of total), chondroitin sulfate A and/or C (10%-20%of total) and little or no hyaluronic acid (0%-5% of total). This pattern of glycosaminoglycan formation differed significantly from that exhibited by isologous skin fibroblasts cultured under identical conditions. Dermal fibroblasts synthesize and secrete primarily hyaluronic acid (50%-60% of total) with lesser amounts of dermatan sulfate (10%-20% of total) and chondroitin sulfate A and/or C (10%-20% of total). These results indicate that differences exist in proteoglycan metabolism between these two connective tissue-producing cells in vitro, and suggest that the observed pattern of in vitro glycosaminoglycan synthesis by primate arterial smooth muscle cells may be characteristic for this cell type and not a general response to conditions of cell culture.

ANP signaling inhibits TGF-β-induced Smad2 and Smad3 nuclear translocation and extracellular matrix expression in rat pulmonary arterial smooth muscle cells

2007 ◽  
Vol 102 (1) ◽  
pp. 390-398 ◽  
Author(s):  
Peng Li ◽  
Suzanne Oparil ◽  
Lea Novak ◽  
Xu Cao ◽  
Weibin Shi ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Nuclear Translocation ◽  
Muscle Cells ◽  
Arterial Smooth Muscle ◽  
Pulmonary Arterial ◽  
Arterial Smooth Muscle Cells ◽  
Pulmonary Arterial Smooth Muscle ◽  
Tgf Β1

Atrial natriuretic peptide (ANP) and transforming growth factor (TGF)-β play important counterregulatory roles in pulmonary vascular adaptation to chronic hypoxia. To define the molecular mechanism of this important interaction, we tested whether ANP-cGMP-protein kinase G (PKG) signaling inhibits TGF-β1-induced extracellular matrix (ECM) expression and defined the specific site(s) at which this molecular merging of signaling pathways occurs. Rat pulmonary arterial smooth muscle cells (PASMCs) were treated with ANP (1 μM) or cGMP (1 mM) with or without pretreatment with PKG inhibitors KT-5823 (1 μM) or Rp-8-bromo-cGMP (Rp-8-Br-cGMP 50 μM), then exposed to TGF-β1 (1 ng/ml) for 5–360 min (for pSmad nuclear translocation and protein analysis) or 24 h (for ECM mRNA expression). Nuclear translocation of pSmad2 and pSmad3 was assessed by fluorescent confocal microscopy. ANP and cGMP inhibited TGF-β1-induced pSmad2 and pSmad3 nuclear translocation and expression of periostin, osteopontin, and plasminogen activator inhibitor-1 mRNA and protein, but not TGF-β1-induced phosphorylation of Smad2 and Smad3. KT-5823 and Rp-8-Br-cGMP blocked ANP/cGMP-induced activation of PKG and inhibition of TGF-β1-stimulated nuclear translocation of pSmad2 and pSmad3 in PASMCs. These results reveal for the first time a precise site at which ANP-cGMP-PKG signaling exerts its antifibrogenic effect on the profibrogenic TGF-β1 signaling pathway: by blocking TGF-β1-induced pSmad2 and pSmad3 nuclear translocation and ECM expression in PASMCs. Blocking nuclear translocation and subsequent binding of pSmad2 and pSmad3 to TGF-β-Smad response elements in ECM genes may be responsible for the inhibitory effects of ANP on TGF-β-induced expression of ECM molecules.

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