Norepinephrine and ANG II stimulate secretion of TGF-beta by neonatal rat cardiac fibroblasts in vitro

1995 ◽  
Vol 268 (4) ◽  
pp. C910-C917 ◽  
Author(s):  
S. A. Fisher ◽  
M. Absher
Keyword(s):  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Cardiac Fibroblasts ◽  
Neonatal Rat ◽  
Heart Cells ◽  
Ang Ii ◽  
Tgf Beta ◽  
Hypertrophic Growth ◽  
Beta 2

Transforming growth factor-beta (TGF-beta) is a ubiquitous growth-regulating protein that is capable of influencing the growth and function of heart cells in vitro. To better understand the role TGF-beta might play as a paracrine mediator of cardiac hypertrophy, the expression, secretion, and growth effects of TGF-beta were examined. Neonatal cardiac fibroblasts in vitro secreted latent TGF-beta 1 and TGF-beta 2 as high as 15 ng/10(6) cells. Angiotensin II (ANG II) and norepinephrine (NE) each augmented up to threefold the expression and secretion of latent TGF-beta 1 and TGF-beta 2 and also induced a shift in isoform predominance from beta 1 to beta 2. Each agent individually produced hypertrophic growth of neonatal cardiocytes and hyperplastic growth of cardiac fibroblasts. Paradoxically, the combination of NE and ANG II at intermediate and high concentrations resulted in less TGF-beta secretion (compared with either agent alone) and in hypertrophic growth of fibroblasts. These results suggest that the growth-promoting effects of ANG II and NE may in part be mediated via a paracrine stimulation of TGF-beta secretion.

Trophic effects of angiotensin II on neonatal rat cardiac myocytes are mediated by cardiac fibroblasts

1995 ◽  
Vol 269 (3) ◽  
pp. E426-E437 ◽  
Author(s):  
N. N. Kim ◽  
F. J. Villarreal ◽  
M. P. Printz ◽  
A. A. Lee ◽  
W. H. Dillmann
Keyword(s):  
Angiotensin Ii ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Specific Binding ◽  
Cardiac Fibroblasts ◽  
Critical Role ◽  
Neonatal Rat ◽  
Ang Ii ◽  
Tgf Beta ◽  
Trophic Effects

Cultured neonatal rat cardiac fibroblasts (NF) and myocytes (NM) were used to examine the distribution of angiotensin II (ANG II) receptors and the potential role of NF in mediating the trophic response to ANG II in the heart. In NM preparations cultured for 2-5 days, specific binding to 125I-ANG II was < 10% of the specific binding in cultured NF. Binding assays, immunocytochemistry, and autoradiography in NM cultured for > 5 days identified two populations of cells, one with fibroblast-like morphology and high density of ANG II receptors and another with low binding, comparable to NM cultures at day 5 or earlier. Conditioned medium (CM) from untreated NF increased cell surface area and net [3H]leucine (Leu) incorporation 1.4-fold in NM. CM from ANG II-treated NF enhanced [3H]Leu incorporation 2.2-fold in NM. This potentiating effect of ANG II was inhibited by losartan and was absent when ANG II was added directly to NM. In addition, studies using antibodies and bioassay for transforming growth factor-beta 1 (TGF-beta 1) suggested that TGF-beta 1 does not mediate the trophic effects of ANG II on NM. We conclude that ANG II receptors are localized predominantly on NF and that ANG II can indirectly stimulate hypertrophy of NM by stimulating NF to produce a transferrable factor(s). These data suggest that cardiac fibroblasts may play a critical role in mediating the hypertrophic response to ANG II in the rat heart.

scholarly journals Transforming growth factor-beta stimulates collagen VII expression by cutaneous cells in vitro.

1992 ◽  
Vol 117 (3) ◽  
pp. 679-685 ◽  
Author(s):  
A König ◽  
L Bruckner-Tuderman
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Human Keratinocytes ◽  
Steady Increase ◽  
Tgf Beta ◽  
Growth Factor Beta ◽  
Beta 2 ◽  
Collagen Vii

Collagen VII, the major component of cutaneous anchoring fibrils is expressed at a low level by normal human keratinocytes and fibroblasts in vitro. In cocultures of these two cell types, signals from fibroblasts enhance expression of collagen VII by keratinocytes and vice versa. In this study, the effects of a possible mediator of such a stimulation, transforming growth factor-beta (TGF-beta), were investigated. Its effect on the expression and deposition of the highly insoluble collagen VII was assessed in a semiquantitative manner by a newly developed enzyme-linked immunoassay which is based on immunoblotting. In keratinocyte monocultures, 0.5-20 ng/ml of TGF-beta 2 induced a dose-dependent stimulation of collagen VII expression as measured per microgram of DNA. The maximal enhancement was about sevenfold compared to controls. The effect of TGF-beta 2 was observed already after 12 h, with a steady increase at least up to 3 d. As previous studies have implicated, untreated cocultures of keratinocytes and fibroblasts exhibited a higher basic level of collagen VII expression, which could be further stimulated about twofold by TGF-beta 2. Fibroblasts alone synthesized very minor quantities of collagen VII and could be only weakly stimulated by TGF-beta 2. This growth factor seems a specific enhancer of collagen VII since the expression of laminin, collagen IV, as well as total protein was increased to a much lesser extent. Our data suggest that TGF-beta may be an important mediator of epithelial-mesenchymal interactions and may regulate the synthesis of the anchoring fibrils at the skin basement membrane zone.

Growth-factor-related proteins that are inducers in early amphibian development may mediate similar steps in amniote (bird) embryogenesis

Development ◽  
1991 ◽  
Vol 111 (1) ◽  
pp. 197-212
Author(s):  
J. Cooke ◽  
A. Wong
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Normal Development ◽  
Tgf Beta ◽  
A Cell ◽  
Growth Factor Beta ◽  
Beta 2 ◽  
Related Proteins

Xenopus and murine activin A homologues (XTC-MIF and WEHI-MIF) and Xenopus and bovine basic fibroblast growth factor (bFGFs) are potent inducers of mesodermal and endodermal pathways of development in amphibian blastular animal cap cells. Porcine transforming growth factor beta 2 (TGF beta 2) is a weaker inducer in the same assay but human platelet-derived growth factor (PDGF) is inactive. We have assayed these factors for evidence of homologous effects in bird development. Unlike amphibians, bird embryos never exhibit a clean segregation of a cell layer that has a uniform specification when uninduced, and can be cultured in isolation as an assay after exposure to soluble factors. We have therefore performed less direct experiments, of three types. We have briefly cultured early chick epiblast cells with and without factors and then assayed their capacity to attach and spread upon fibronectin, in comparison with young streak and substreak hypoblast cells. We have asked whether similar microculture with factors alters the ability of quail epiblast cells to disrupt morphogenesis, and to integrate into the structure, of host chick blastoderms into which they are seeded. Finally, whole early chick blastoderms have been preincubated with or without factors for a brief period before setting them up to develop in vitro under circumstances usually permitting successful formation of axial pattern. Strong effects of the activin-like factors, of bFGF and of TGF beta 2 were seen in all three procedures, while PDGF was essentially inactive. In epiblast cells, effective factors at picomolar concentrations induced stable spreading upon fibronectin, and a capacity to adhere and spread upon basal epiblast surface and prevent morphogenesis in host blastoderms. Preincubation of whole early blastoderms with these factors led to characteristic deviation from normal development over the subsequent 24 h. We therefore suggest that peptides from the particular families that are active as inducers in amphibian blastula ectoderm may mediate homologous or closely related steps in respecification throughout vertebrates.

Dominant negative mutants of transforming growth factor-beta 1 inhibit the secretion of different transforming growth factor-beta isoforms

1992 ◽  
Vol 12 (4) ◽  
pp. 1674-1679
Author(s):  
A R Lopez ◽  
J Cook ◽  
P L Deininger ◽  
R Derynck
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Mammalian Cells ◽  
Transforming Growth Factor ◽  
Dominant Negative ◽  
Tgf Beta ◽  
Growth Factor Beta ◽  
Beta 2

Transforming growth factor-beta (TGF-beta) is a secreted polypeptide factor that is thought to play a major role in the regulation of proliferation of many cell types and various differentiation processes. Several related isoforms have been structurally characterized, three of which, TGF-beta 1, -beta 2, and -beta 3, have been detected in mammalian cells and tissues. Each TGF-beta form is a homodimer of a 112-amino-acid polypeptide which is encoded as a larger polypeptide precursor. We have introduced several mutations in the TGF-beta 1 precursor domain, resulting in an inhibition of TGF-beta 1 secretion. Coexpression of these mutants with wild-type TGF-beta 1, -beta 2, and -beta 3 results in a competitive and specific inhibition of the secretion of different TFG-beta forms, indicating that these mutated versions act as dominant negative mutants for TGF-beta secretion. Overexpression of dominant negative mutants can thus be used to abolish endogenous secretion of TGF-beta and structurally related family members, both in vitro and in vivo, and to probe in this way the physiological functions of the members of the TGF-beta superfamily.

scholarly journals Dominant negative mutants of transforming growth factor-beta 1 inhibit the secretion of different transforming growth factor-beta isoforms.

1992 ◽  
Vol 12 (4) ◽  
pp. 1674-1679 ◽  
Author(s):  
A R Lopez ◽  
J Cook ◽  
P L Deininger ◽  
R Derynck
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Mammalian Cells ◽  
Transforming Growth Factor ◽  
Dominant Negative ◽  
Tgf Beta ◽  
Growth Factor Beta ◽  
Beta 2

Transforming growth factor-beta (TGF-beta) is a secreted polypeptide factor that is thought to play a major role in the regulation of proliferation of many cell types and various differentiation processes. Several related isoforms have been structurally characterized, three of which, TGF-beta 1, -beta 2, and -beta 3, have been detected in mammalian cells and tissues. Each TGF-beta form is a homodimer of a 112-amino-acid polypeptide which is encoded as a larger polypeptide precursor. We have introduced several mutations in the TGF-beta 1 precursor domain, resulting in an inhibition of TGF-beta 1 secretion. Coexpression of these mutants with wild-type TGF-beta 1, -beta 2, and -beta 3 results in a competitive and specific inhibition of the secretion of different TFG-beta forms, indicating that these mutated versions act as dominant negative mutants for TGF-beta secretion. Overexpression of dominant negative mutants can thus be used to abolish endogenous secretion of TGF-beta and structurally related family members, both in vitro and in vivo, and to probe in this way the physiological functions of the members of the TGF-beta superfamily.

scholarly journals Transforming growth factor beta selectively inhibits normal and leukemic human bone marrow cell growth in vitro

Blood ◽  
1988 ◽  
Vol 72 (5) ◽  
pp. 1504-1511 ◽  
Author(s):  
GK Sing ◽  
JR Keller ◽  
LR Ellingsworth ◽  
FW Ruscetti
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Bone Marrow Cells ◽  
Tgf Beta ◽  
Gm Csf ◽  
Growth Factor Beta ◽  
Beta 2

Abstract The effects of transforming growth factor beta 1 or beta 2 (TGF-beta 1 or -beta 2) on the in vitro proliferation and differentiation of normal and malignant human hematopoietic cells were studied. Both forms of TGF- beta suppressed both the normal cellular proliferation and colony formation induced by recombinant human interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF). In the presence of GM-CSF or IL-3, optimal concentrations of TGF-beta (400 pmol/L) inhibited colony formation by erythroid (BFU-E), multipotential (CFU-GEMM), and granulocyte-macrophage (CFU-GM) progenitor cells by 90% to 100%, whereas granulocyte or monocyte cluster formation was not inhibited. In contrast, neither form of TGF-beta had any effect on G- CSF-induced hematopoiesis. The suppressive action appeared to be mediated directly by TGF-beta since antiproliferative responses were also observed in accessory cell-depleted bone marrow cells. In contrast to normal bone marrow cells, both GM- and G-CSF-induced proliferation of cells from patients with chronic myelogenous leukemia were suppressed in a dose-dependent manner by TGF-beta. Differential effects of TGF-beta on the proliferation of established leukemic lines were also observed since most cell lines of myelomonocytic nature studied were strongly inhibited where erythroid cell lines were either insensitive or poorly inhibited by TGF-beta. These results suggest that TGF-beta is an important modulator of human hematopoiesis that selectively regulates the growth of less mature hematopoietic cell populations with a high proliferative capacity as opposed to more differentiated cells, which are not affected by TGF-beta.

scholarly journals Transforming growth factor beta selectively inhibits normal and leukemic human bone marrow cell growth in vitro

Blood ◽  
1988 ◽  
Vol 72 (5) ◽  
pp. 1504-1511 ◽  
Author(s):  
GK Sing ◽  
JR Keller ◽  
LR Ellingsworth ◽  
FW Ruscetti
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Bone Marrow Cells ◽  
Tgf Beta ◽  
Gm Csf ◽  
Growth Factor Beta ◽  
Beta 2

The effects of transforming growth factor beta 1 or beta 2 (TGF-beta 1 or -beta 2) on the in vitro proliferation and differentiation of normal and malignant human hematopoietic cells were studied. Both forms of TGF- beta suppressed both the normal cellular proliferation and colony formation induced by recombinant human interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF). In the presence of GM-CSF or IL-3, optimal concentrations of TGF-beta (400 pmol/L) inhibited colony formation by erythroid (BFU-E), multipotential (CFU-GEMM), and granulocyte-macrophage (CFU-GM) progenitor cells by 90% to 100%, whereas granulocyte or monocyte cluster formation was not inhibited. In contrast, neither form of TGF-beta had any effect on G- CSF-induced hematopoiesis. The suppressive action appeared to be mediated directly by TGF-beta since antiproliferative responses were also observed in accessory cell-depleted bone marrow cells. In contrast to normal bone marrow cells, both GM- and G-CSF-induced proliferation of cells from patients with chronic myelogenous leukemia were suppressed in a dose-dependent manner by TGF-beta. Differential effects of TGF-beta on the proliferation of established leukemic lines were also observed since most cell lines of myelomonocytic nature studied were strongly inhibited where erythroid cell lines were either insensitive or poorly inhibited by TGF-beta. These results suggest that TGF-beta is an important modulator of human hematopoiesis that selectively regulates the growth of less mature hematopoietic cell populations with a high proliferative capacity as opposed to more differentiated cells, which are not affected by TGF-beta.

scholarly journals Transforming growth factor beta suppresses human immunodeficiency virus expression and replication in infected cells of the monocyte/macrophage lineage.

1991 ◽  
Vol 173 (3) ◽  
pp. 589-597 ◽  
Author(s):  
G Poli ◽  
A L Kinter ◽  
J S Justement ◽  
P Bressler ◽  
J H Kehrl ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
T Cell ◽  
Growth Factor ◽  
Cell Line ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Tgf Beta ◽  
Immunodeficiency Virus ◽  
Growth Factor Beta

The pleiotropic immunoregulatory cytokine transforming growth factor beta (TGF-beta) potently suppresses production of the human immunodeficiency virus (HIV), the causative agent of the acquired immunodeficiency syndrome, in the chronically infected promonocytic cell line U1. TGF-beta significantly (50-90%) inhibited HIV reverse transcriptase production and synthesis of viral proteins in U1 cells stimulated with phorbol myristate acetate (PMA) or interleukin 6 (IL-6). Furthermore, TGF-beta suppressed PMA induction of HIV transcription in U1 cells. In contrast, TGF-beta did not significantly affect the expression of HIV induced by tumor necrosis factor alpha (TNF-alpha). These suppressive effects were not mediated via the induction of interferon alpha (IFN-alpha). TGF-beta also suppressed HIV replication in primary monocyte-derived macrophages infected in vitro, both in the absence of exogenous cytokines and in IL-6-stimulated cultures. In contrast, no significant effects of TGF-beta were observed in either a chronically infected T cell line (ACH-2) or in primary T cell blasts infected in vitro. Therefore, TGF-beta may play a potentially important role as a negative regulator of HIV expression in infected monocytes or tissue macrophages in infected individuals.

Abstract 782: Fra2 Mediates Oxygen-induced TGFbeta-1 mRNA Expression In Adult Cardiac Fibroblasts: Significance In Myocardial Fibrosis Following Ischemia-reperfusion Injury

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Sashwati Roy ◽  
Savita Khanna ◽  
Chandan K Sen
Keyword(s):  
Mrna Expression ◽  
Transforming Growth Factor Beta ◽  
Cardiac Fibrosis ◽  
Transforming Growth Factor ◽  
Ischemia Reperfusion Injury ◽  
Cardiac Fibroblasts ◽  
Ischemia Reperfusion ◽  
Luciferase Reporter ◽  
Mrna Levels

Background . Transforming growth factor beta-1 (TGFbeta-1) is a key cytokine implicated in the development of cardiac fibrosis following ischemia-reperfusion (IR) injury. The profibrotic effects of TGFbeta-1 are primarily attributable to the differentiation of cardiac fibroblasts (CF) to myofibroblasts. Previously, we have reported perceived hyperoxia (Circ Res 92:264 –71), sub-lethal reoxygenation shock during IR, induces differentiation of CF to myofibroblasts at the infarct site. The mechanisms underlying oxygen-sensitive induction of TGFbeta-1 mRNA remain to be characterized. Hypothesis . Fra2 mediates oxygen-induced TGFbeta-1 mRNA expression in adult cardiac fibroblasts. Methods. TGFbeta-1 mRNA expression in infarct tissue was investigated in an IR injury model. The left anterior descending coronary artery of mice was transiently occluded for 60 minutes followed by reperfusion to induce IR injury. Spatially resolved infarct and non-infarct tissues were collected at 0, 1, 3, 5, and 7 days post-IR using laser capture microdissection. TGFbeta-1 mRNA levels were measured using real-time PCR. To investigate the role of oxygen in the regulation of TGFbeta-1, we used our previously reported model of perceived hyperoxia where CF (from 5wks old mice) after isolation were cultured at 5%O 2 (physiological pO 2 ) followed by transferring them to 20%O 2 to induce hyperoxic insult. Results & Conclusions. In vivo, a significant increase (p<0.01; n=5) in TGFbeta-1 mRNA was observed at the infarct site already at day 1 post-IR. The levels continued to increase until day 7 post-IR. In vitro, exposure of CF to 20%O 2 hyperoxic insult induced TGFbeta-1 mRNA (p<0.001; n=4) and protein (p<0.01; n=4) expression. Using a TGFbeta-1 promoter-luciferase reporter and DNA binding assays, we collected first evidence that AP-1 and its component Fra2 as major mediators of oxygen-induced TGFbeta-1 expression. Exposure to 20%O 2 resulted in increased localization of Fra2 in nucleus. siRNA-dependent Fra-2 knock-down completely abrogated oxygen-induced TGFbeta1 expression. In conclusion, this study presents first evidence that Fra-2 is involved in inducible TGFbeta1 expression in CF. Fra2 was noted as being central in regulating oxygen-induced TGFbeta-1 expression.s

Neutralisation of TGF-beta 1 and TGF-beta 2 or exogenous addition of TGF-beta 3 to cutaneous rat wounds reduces scarring

1995 ◽  
Vol 108 (3) ◽  
pp. 985-1002 ◽  
Author(s):  
M. Shah ◽  
D.M. Foreman ◽  
M.W. Ferguson
Keyword(s):  
Wound Healing ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Collagen I ◽  
Tgf Beta ◽  
Early Stages ◽  
Collagen Iii ◽  
Exogenous Addition ◽  
Growth Factor Beta ◽  
Beta 2

Exogenous addition of neutralising antibody to transforming growth factor-beta 1,2 to cutaneous wounds in adult rodents reduces scarring. Three isoforms of transforming growth factor-beta (1, 2 and 3) have been identified in mammals. We investigated the isoform/isoforms of TGF-beta responsible for cutaneous scarring by: (i) reducing specific endogenous TGF-beta isoforms by exogenous injection of isoform specific neutralising antibodies; and (ii) increasing the level of specific TGF-beta isoforms by exogenous infiltration into the wound margins. Exogenous addition of neutralising antibody to TGF-beta 1 plus neutralising antibody to TGF-beta 2 reduced the monocyte and macrophage profile, neovascularisation, fibronectin, collagen III and collagen I deposition in the early stages of wound healing compared to control wounds. Treatment with neutralising antibodies to TGF-betas 1 and 2 markedly improved the architecture of the neodermis to resemble that of normal dermis and reduced scarring while the control wounds healed with scar formation. Exogenous addition of neutralising antibody to TGF-beta 1 alone also reduced the monocyte and macrophage profile, fibronectin, collagen III and collagen I deposition compared to control wounds. However, treatment with neutralising antibody to TGF-beta 1 alone only marginally reduced scarring. By contrast, wounds treated with neutralising antibody to TGF-beta 2 alone did not differ from control wounds. Interestingly, exogenous addition of the TGF-beta 3 peptide also reduced the monocyte and macrophage profile, fibronectin, collagen I and collagen III deposition in the early stages of wound healing and markedly improved the architecture of the neodermis and reduced scarring. By contrast, wounds treated with either TGF-beta 1 or with TGF-beta 2 had more extracellular matrix deposition in the early stages of wound healing but did not differ from control wounds in the final quality of scarring. This study clearly demonstrates isoform specific differences in the role of TGF-betas in wound healing and cutaneous scarring. TGF-beta 1 and TGF-beta 2 are implicated in cutaneous scarring. This study also suggests a novel therapeutic use of exogenous recombinant, TGF-beta 3 as an anti-scarring agent.

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