Collagen type I enhances endothelin-mediated contraction and induces nonproliferating phenotype in mesangial cells

1996 ◽  
Vol 270 (6) ◽  
pp. F960-F970 ◽  
Author(s):  
T. Miralem ◽  
C. I. Whiteside ◽  
D. M. Templeton
Keyword(s):  
Light Chain ◽  
Myosin Light Chain ◽  
Collagen Type ◽  
Mesangial Cells ◽  
Collagen Gel ◽  
Collagen Type I ◽  
Cell Number ◽  
Type I ◽  
Total Dna ◽  
Glomerular Extracellular Matrix

Accumulation of glomerular extracellular matrix is a characteristic accompaniment of mesangial cell proliferation in progressive renal disease. We examined how growth on several matrices affected the proliferative phenotype of cultured rat mesangial cells. Compared with growth on plastic, Matrigel, or mesangial matrix, collagen type I caused a decreased cell number at 72 h, decreased total DNA per culture, and a decrease in the incorporation of [3H]thymidine during S phase in cells released from quiescence. These antiproliferative and antimitogenic effects of collagen type I required growth on a collagen gel; soluble collagen or collagen fragments were without effect. Because a number of agents elicit both proliferative and contractile responses in mesangial cells, we examined the effect of growth on collagen on contractility. Compared with plastic, cells grown on collagen type I were more contractile, showed a higher Ca2+ signal in response to endothelin, and responded to endothelin with a more rapid myosin light-chain kinase-dependent phosphorylation of myosin light chain. We conclude that growth on a collagen type I gel uncouples contractility from a proliferative response in mesangial cells, suppressing proliferation while enhancing contraction and Ca2+ signaling in response to endothelin.

scholarly journals Stimulation of beta1 integrins on fibroblasts induces PDGF independent tyrosine phosphorylation of PDGF beta-receptors.

1996 ◽  
Vol 132 (4) ◽  
pp. 741-752 ◽  
Author(s):  
C Sundberg ◽  
K Rubin
Keyword(s):  
Cell Adhesion ◽  
Tyrosine Phosphorylation ◽  
Collagen Type ◽  
Collagen Gel ◽  
Collagen Type I ◽  
Receptor Activation ◽  
Type I ◽  
Integrin Subunit ◽  
Beta Receptor ◽  
Beta Receptors

We report that integrin-mediated signaling induces a rapid and transient tyrosine phosphorylation of platelet-derived growth factor (PDGF) beta-receptors in human diploid foreskin AG 1518 fibroblasts. A transient tyrosine phosphorylation of PDGF beta-receptors was evident one and two hours after cells had been plated on collagen type I and fibronectin, as well as on immobilized anti-integrin subunit IgG, but not on poly-L-lysine. In contrast EGF or PDGF alpha-receptors were not phosphorylated on tyrosine residues under these conditions. Tyrosine phosphorylation of PDGF beta-receptors induced by plating on collagen type I was inhibited by cytochalasin D and herbimycin A, unaffected by cycloheximide and enhanced by orthovanadate. Furthermore, a transient phosphorylation of PDGF beta-receptors occurred when AG 518 fibroblasts were cultured in three-dimensional collagen lattices or exposed to external strain exerted through centrifugation. The latter effect was evident already after two minutes. Clustering of cell surface beta1 integrins led to PDGF beta-receptor phosphorylation both in suspended and firmly attached AG 1518 fibroblasts. Plating of cells on collagen type I, fibronectin, and anti-beta1-integrin IgG resulted in the formation of PDGF beta-receptor aggregates as detected by immunofluorescence. Suramin or anti-PDGF-BB IgG had no effect on the plating-induced tyrosine phosphorylation of PDGF beta-receptors. PDGF-B chain mRNA, or protein, were not detected in AG 1518 fibroblasts. Our data suggest that a ligand-independent PDGF beta-receptor activation during cell adhesion and early phases of cell spreading is involved in integrin-mediated signaling in fibroblasts, and constitutes parts of a mechanism for cells to respond during the dynamic phases of externally applied tension as well as fibroblast-mediated tension during cell adhesion and collagen gel contraction.

Changes in extracellular matrix composition regulate cyclooxygenase-2 expression in human mesangial cells

2011 ◽  
Vol 300 (4) ◽  
pp. C907-C918 ◽  
Author(s):  
Matilde Alique ◽  
Laura Calleros ◽  
Alicia Luengo ◽  
Mercedes Griera ◽  
Miguel Ángel Iñiguez ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Collagen Type ◽  
Mesangial Cells ◽  
Collagen Type I ◽  
Collagen I ◽  
Type I ◽  
Camp Response Element ◽  
Response Element ◽  
Human Mesangial Cells ◽  
Cox 2

Glomerular diseases are characterized by a sustained synthesis and accumulation of abnormal extracellular matrix proteins, such as collagen type I. The extracellular matrix transmits information to cells through interactions with membrane components, which directly activate many intracellular signaling events. Moreover, accumulating evidence suggests that eicosanoids derived from cyclooxygenase (COX)-2 participate in a number of pathological processes in immune-mediated renal diseases, and it is known that protein kinase B (AKT) may act through different transcription factors in the regulation of the COX-2 promoter. The present results show that progressive accumulation of collagen I in the extracellular medium induces a significant increase of COX-2 expression in human mesangial cells, resulting in an enhancement in PGE2 production. COX-2 overexpression is due to increased COX-2 mRNA levels. The study of the mechanism implicated in COX-2 upregulation by collagen I showed focal adhesion kinase (FAK) activation. Furthermore, we observed that the activation of the phosphatidylinositol 3-kinase (PI3K)/AKT pathway by collagen I and collagen I-induced COX-2 overexpression was abolished by PI3K and AKT inhibitors. Additionally, we showed that the cAMP response element (CRE) transcription factor is implicated. Finally, we studied COX-2 expression in an animal model, NG-nitro-l-arginine methyl ester hypertensive rats. In renal tissue and vascular walls, COX-2 and collagen type I content were upregulated. In summary, our results provide evidence that collagen type I increases COX-2 expression via the FAK/PI3K/AKT/cAMP response element binding protein signaling pathway.

Bacterial Endotoxin Inhibits Migration, Attachment, and Orientation of Human Gingival Fibroblasts in vitro and Delays Collagen Gel Contraction

1987 ◽  
Vol 66 (9) ◽  
pp. 1449-1455 ◽  
Author(s):  
S. Pitaru ◽  
M. Soldinger ◽  
D. Madgar ◽  
Z. Metzger
Keyword(s):  
Collagen Type ◽  
Cell Attachment ◽  
Collagen Gel ◽  
Collagen Type I ◽  
Human Gingival Fibroblasts ◽  
Bacterial Endotoxin ◽  
Type I ◽  
Gingival Fibroblasts ◽  
Collagen Gels

The purpose of this study was to assess the effect of endotoxin adsorbed to dental surfaces and to collagen type I on the migration, attachment, and orientation of human gingival fibroblasts (HGF). Transversely cut porcine tooth root slices (RS), 200 μm thick, were prepared. Half of the RS obtained were partially demineralized in EDTA. Half of the demineralized and non-demineralized RS were incubated with 400 μg/mL of endotoxin for 24 hr, whereas the other half were maintained in PBS and served as controls. Experimental and control RS were placed on confluent layers of HFG and cultured for six days. Cell migration toward and cell attachment to the periphery of the RS and the formation of oriented cell sheets were assessed by means of photographic techniques. Additionally, six-day-old cultures were fixed and processed for SEM observation. In separate experiments, the effect of endotoxin on cell attachment to collagen type I and on contraction of three-dimensional collagen gels was assessed. It was found that: (i) bacterial endotoxin inhibited migration and attachment of HGF to both demineralized and non-demineralized cementum and interfered with the development of oriented cellular structure ; (ii) the inhibitory effect was significantly more pronounced for non-demineralized than for demineralized cementum; (iii) the morphology of HGF attached to endotoxin-treated dental surfaces was altered compared with that of their controls; and (iv) bacterial endotoxin inhibited cell attachment to collagen type I and delayed the contraction of collagen gel.

Effects of Controlled Released BMP-7 on Markers of Inflammation and Degradation During the Cultivation of Human Osteoarthritic Chondrocytes

2010 ◽  
Vol 26 (4) ◽  
pp. 419-433 ◽  
Author(s):  
Karsten Gavenis ◽  
Thomas Pufe ◽  
Lars Ove Brandenburg ◽  
Katharina Schiffl ◽  
Bernhard Schmidt-Rohlfing
Keyword(s):  
Gene Expression ◽  
Collagen Type ◽  
Articular Chondrocytes ◽  
Collagen Gel ◽  
Collagen Type I ◽  
Type I ◽  
Matrix Synthesis ◽  
Gene Expressions ◽  
Markers Of Inflammation ◽  
Osteoarthritic Chondrocytes

The aim of the present study is to investigate the effects of BMP-7 released from polylactide microspheres on the appearance of various catabolic and inflammatory cytokines secreted by osteoarthritic chondrocytes cultivated in a collagen gel. Articular chondrocytes of 15 patients suffering from osteoarthritis are transferred to a collagen type-I gel. Additionally, BMP-7 encapsulated into polylactide microspheres (50 ng BMP-7/mL gel) is added. After 14 days, gene expression and protein appearance of various genes involved in matrix turnover and inflammation are investigated by immunohistochemical staining and RT-PCR and compared to untreated controls. TNF-α, MMP-13, IL-6, IL-1β, and VEGF gene expressions are decreased in the treatment group. In contrast, BMP-7-induced matrix synthesis is not affected, leaving collagen type-II (Col-II) gene expression to be elevated, while collagen type-I (Col-I) is decreased. In summary, controlled release of low concentrated BMP-7 from polylactide microspheres leads to a decrease in gene expression of the investigated inflammation and matrix degradation markers whereas matrix synthesis is induced.

scholarly journals Biological Effects of Lentivirus-Mediated shRNA Targeting Collagen Type I on the Mesangial Cells of Rats

Renal Failure ◽  
2011 ◽  
Vol 33 (3) ◽  
pp. 334-340 ◽  
Author(s):  
Chunhua Zhou ◽  
Yi Shan ◽  
Haidan Zhao ◽  
Ping He
Keyword(s):  
Collagen Type ◽  
Mesangial Cells ◽  
Biological Effects ◽  
Collagen Type I ◽  
Type I

TGF-β-regulated collagen type I accumulation: role of Src-based signals

2007 ◽  
Vol 292 (4) ◽  
pp. C1361-C1369 ◽  
Author(s):  
Rangnath Mishra ◽  
Ling Zhu ◽  
Richard L. Eckert ◽  
Michael S. Simonson
Keyword(s):  
Type I Collagen ◽  
Collagen Type ◽  
Transforming Growth Factor ◽  
Mesangial Cells ◽  
Collagen Type I ◽  
Dominant Negative ◽  
Dominant Negative Mutant ◽  
Type I ◽  
Small Interference Rna ◽  
Collagen Secretion

Transforming growth factor-β (TGF-β) stimulates myofibroblast transdifferentiation, leading to type I collagen accumulation and fibrosis. We investigated the function of Src in TGF-β-induced collagen I accumulation. In human mesangial cells, PTyr416 Src (activated Src) was 3.3-fold higher in TGF-β-treated cells than in controls. Src activation by TGF-β was blocked by rottlerin and by a dominant negative mutant of protein kinase Cδ (PKCδ), showing that TGF-β activates Src by a PKCδ-based mechanism. Pharmacological inhibitors and a dominant negative Src mutant prevented the increase in collagen type I secretion in cells exposed to TGF-β. Similarly, on-target Src small interference RNA (siRNA) prevented type I collagen secretion in response to TGF-β, but off-target siRNA complexes had no effect. It is well established in mesangial cells that upregulation of type I collagen by TGF-β requires extracellular signal-regulated kinase 1/2 (ERK1/2), and we found that activation of ERK1/2 by TGF-β requires Src. In conclusion, these results suggest that stimulation of collagen type I secretion by TGF-β requires a PKCδ-Src-ERK1/2 signaling motif.

scholarly journals Inactivation of kinase cascades in mesangial cells grown on collagen type I

1998 ◽  
Vol 275 (4) ◽  
pp. F585-F594 ◽  
Author(s):  
Tiho Miralem ◽  
Douglas M. Templeton
Keyword(s):  
Collagen Type ◽  
Mesangial Cell ◽  
Mesangial Cells ◽  
Collagen Type I ◽  
Signaling Cascades ◽  
Renal Diseases ◽  
Type I ◽  
Tyrosine Residues ◽  
Kinase C ◽  
Dependent Protein

Growth on collagen type I gels is known to suppress the mitogenic responsiveness of mesangial cells. Because these cells proliferate in some renal diseases and themselves synthesize collagen type I, we examined the influence of growth on collagen upon several kinase signaling cascades involved in mesangial cell proliferation. Quiescent mesangial cells grown on collagen type I and then stimulated with serum showed a markedly diminished induction of the protooncogene c- fos, compared with their counterparts on plastic or fibronectin. This effect was accompanied by decreased activation of mitogen-activated (Erk family) and Ca2+/calmodulin-dependent protein kinases. Cells on collagen showed lower basal protein kinase C (PKC) activity and diminished levels of PKC-α and -ζ isoforms. Global phosphorylation of tyrosine residues was diminished on collagen, and tyrosine phosphorylation of Erk and focal adhesion kinase in response to serum was not detected, in contrast to cells on plastic. We conclude that attachment of mesangial cells to collagen type I results in a broad suppression of protein phosphorylation that is reflected in diminished induction of the c- fos gene and probably underlies the conversion of cultured mesangial cells to a nonproliferative phenotype.

TNF-α, but not IFN-γ, regulates CCN2 (CTGF), collagen type I, and proliferation in mesangial cells: possible roles in the progression of renal fibrosis

2007 ◽  
Vol 293 (1) ◽  
pp. F157-F165 ◽  
Author(s):  
Laurinda A. Cooker ◽  
Darryl Peterson ◽  
Joann Rambow ◽  
Melisa L. Riser ◽  
Rebecca E. Riser ◽  
...  
Keyword(s):  
Renal Fibrosis ◽  
Cellular Proliferation ◽  
Collagen Type ◽  
Mesangial Cells ◽  
Collagen Type I ◽  
Term Treatment ◽  
Type I ◽  
Mrna Levels ◽  
Tnf Α ◽  
Ifn Γ

Connective tissue growth factor (CCN2) is a profibrotic factor acting downstream and independently of TGF-β to mediate renal fibrosis. Although inflammation is often involved in the initiation and/or progression of fibrosis, the role of inflammatory cytokines in regulation of glomerular CCN2 expression, cellular proliferation, and extracellular matrix accumulation is unknown. We studied two such cytokines, TNF-α and IFN-γ, for their effects on cultured mesangial cells in the presence or absence of TGF-β, as a model for progressive renal fibrosis. Short-term treatment with TNF-α, like TGF-β, significantly increased secreted CCN2 per cell, but unlike TGF-β inhibited cellular replication. TNF-α combined with TGF-β further increased CCN2 secretion and mRNA levels and reduced proliferation. Surprisingly, however, TNF-α treatment decreased baseline collagen type I protein and mRNA levels and largely blocked their stimulation by TGF-β. Long-term treatment with TGF-β or TNF-α alone no longer increased CCN2 protein levels. However, the combination synergistically increased CCN2. IFN-γ had no effect on either CCN2 or collagen activity and produced a mild inhibition of TGF-β-induced collagen only at a high concentration (500 U/ml). In summary, we report a strong positive regulatory role for TNF-α, but not IFN-γ, in CCN2 production and secretion, including that driven by TGF-β. The stimulation of CCN2 release by TNF-α, unlike TGF-β, is independent of cellular proliferation and not linked to increased collagen type I accumulation. This suggests that the paradigm of TGF-β-driven CCN2 with subsequent collagen production may be overridden by an as yet undefined inhibitory mechanism acting either directly or indirectly on matrix metabolism.

scholarly journals SPARC Regulates the Expression of Collagen Type I and Transforming Growth Factor-β1 in Mesangial Cells

1999 ◽  
Vol 274 (45) ◽  
pp. 32145-32152 ◽  
Author(s):  
Aleksandar Francki ◽  
Amy D. Bradshaw ◽  
James A. Bassuk ◽  
Chin C. Howe ◽  
William G. Couser ◽  
...  
Keyword(s):  
Growth Factor ◽  
Collagen Type ◽  
Transforming Growth Factor ◽  
Mesangial Cells ◽  
Collagen Type I ◽  
Transforming Growth Factor Β1 ◽  
Type I

scholarly journals Effect of a Direct Thrombin Inhibitor Compared with Dalteparin and Unfractionated Heparin on Human Osteoblasts

2011 ◽  
Vol 5 (1) ◽  
pp. 52-58 ◽  
Author(s):  
Tobias Winkler ◽  
Carsten Perka ◽  
Dörte Matziolis ◽  
Georg Matziolis
Keyword(s):  
Collagen Type ◽  
Collagen Type I ◽  
Cell Number ◽  
Direct Thrombin Inhibitor ◽  
Thrombin Inhibitors ◽  
Thrombin Inhibitor ◽  
Direct Thrombin Inhibitors ◽  
Type I ◽  
Human Osteoblasts

Purpose:Osteoporosis is a relevant problem after long term administration of unfractionated heparin (UFH) and low molecular weight heparin. Melagatran is a representative of a new group of direct thrombin inhibitors with comparable data in the prevention of thromboembolic events after orthopaedic surgery. The aim of ourin vitrostudy was to investigate the effect of a direct thrombin inhibitor compared with dalteparin and UFH on human osteoblasts.Materials and Methods:Melagatran, dalteparin and UFH were added to primary osteoblast cultures in their therapeutic range and two decimal powers below and above. Cell number, protein synthesis, mitochondrial and alkaline phosphatase activity and collagen type I synthesis were evaluated.Results:Melagatran showed the least influence on protein synthesis and cell proliferation with a reduction of cell number to 83.5 ± 9% (p = 0.027) of the control group only in the highest investigated concentration after 15 days of incubation.Mitochondrial and alkaline phosphatase activity and collagen type I synthesis in osteoblasts incubated with melagatran and dalteparin showed similar patterns. UFH showed the most pronounced influence on cellular metabolism.Conclusions:Melagatran showed less inhibitory in vitro effects on human osteoblasts than dalteparin or UFH. The presented study gives first hints that direct thrombin inhibitors may help prevent heparin-induced negative effects on bone metabolism.

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