TGF-β1-mediated fibroblast–myofibroblast terminal differentiation—the role of smad proteins

Evans, Rachel C.; Tian, Ya‐Chung; Steadman, Robert; Phillips, Aled Owain

doi:10.1016/s0014-4827(02)00015-0

Cited by 338 publications

(285 citation statements)

References 23 publications

Supporting

Mentioning

262

Contrasting

Unclassified

Order By: Relevance

“…In addition, fibroblasts transfected with Smad2 had up-regulated α-SMA mRNA expression and altered cell phenotype [Evans et al, 2003]. In our study, active Smad2 could not be detected after 30 h of TGF-β1 treatment in the presence or absence of nicotine.…”

Section: Nicotine Inhibited Tgf-β1-induced P38 Mapk Activitymentioning

confidence: 53%

“…However, treatment of primary neonatal dermal fibroblasts from Smad3 knockout mice with 5 ng/ml TGF-β1 for 4 days induced α-SMA protein at the same level as fibroblasts from wild-type mice [Flanders et al, 2003]. Furthermore, lung fibroblasts transfected with Smad2, but not Smad3 had up-regulated α-SMA mRNA expression and altered cell phenotypes [Evans et al, 2003]. These data suggested that Smad3 was not required for the differentiation of fibroblasts to myofibroblasts.…”

Section: Discussionmentioning

confidence: 94%

See 1 more Smart Citation

Nicotine inhibits myofibroblast differentiation in human gingival fibroblasts

Fang

Svoboda

2005

J of Cellular Biochemistry

View full text Add to dashboard Cite

Cigarette smoking has been suggested as a risk factor for several periodontal diseases. It has also been found that smokers respond less favorably than non-smokers to periodontal therapy. Previous work in our lab has shown that nicotine inhibits human gingival cell migration. Since myofibroblasts play an important role in wound closure, we asked if nicotine affects gingival wound healing process by regulating myofibroblast differentiation. Human gingival fibroblasts (HGFs) from two patients were cultured in 10% fetal bovine serum cell culture medium. Cells were pretreated with different doses of nicotine (0, 0.01, 0.1, and 1 mM) for 2 h, and then incubated with transforming growth factor beta (TGF-β1) (0, 0.25, 0.5, and 1 ng/ml) with or without nicotine for 30 h. The expression level of α-smooth muscle actin (α-SMA), a specific marker for myofibroblasts, was analyzed by Western blots, immunocytochemistry, and real-time polymerase chain reaction (real-time PCR). Phosphorylated p38 mitogen-activated protein kinase (Phospho-p38 MAPK) activity was analyzed by Western blots. TGF-β1 induced an increase of α-SMA protein and mRNA expression, while nicotine (1 mM) inhibited the TGF-β1-induced expression of α-SMA but not β-actin. Nicotine treatment down-regulated TGF-β1-induced p38 MAPK phosphorylation. Our results demonstrated for the first time that nicotine inhibits myofibroblast differentiation in human gingival fibroblasts in vitro; supporting the hypothesis that delayed wound healing in smokers may be due to decreased wound contraction by myofibroblasts. Keywordshuman gingival fibroblasts; myofibroblasts; α smooth muscle actin; TGFβ; p38 MAP kinase Myofibroblasts are contractile cells that share characteristics of fibroblasts and smooth muscle cells [Tomasek et al., 2002]. One of the major features of myofibroblast differentiation is the expression of smooth muscle cell markers. These markers include α-SMA, γ-SMA, smooth muscle 22 (SM22), calponin, smoothelin, meta-vinculin, h-caldesmon [Halayko and Solway, 2001]. Among these, α-SMA has been suggested as the most reliable marker of myofibroblast differentiation [Gabbiani, 2003]. Also, Dr. Masur's lab demonstrated that myofibroblast differentiation was cell density dependent in corneal fibroblasts. They found that the lower the cell density when cells were seeded in culture, the more cells differentiated into myofibroblasts. However, the myofibroblast phenotype was not terminal. The cells lost the myofibroblast

show abstract

Section: Nicotine Inhibited Tgf-β1-induced P38 Mapk Activitymentioning

confidence: 53%

Section: Discussionmentioning

confidence: 94%

Nicotine inhibits myofibroblast differentiation in human gingival fibroblasts

Fang

Svoboda

2005

J of Cellular Biochemistry

View full text Add to dashboard Cite

show abstract

“…lung fibroblasts (27)(28)(29). We examined whether TGFb1 regulates the expression of CCN family members in human lung fibroblasts.…”

Section: Tgf-b1 Up-regulates Ccn1 In Lung Fibroblastsmentioning

confidence: 99%

The matricellular protein CCN1 enhances TGF‐β1/SMAD3‐dependent profibrotic signaling in fibroblasts and contributes to fibrogenic responses to lung injury

Kurundkar

Rangarajan

et al. 2016

FASEB j.

View full text Add to dashboard Cite

Matricellular proteins mediate pleiotropic effects during tissue injury and repair. CCN1 is a matricellular protein that has been implicated in angiogenesis, inflammation, and wound repair. In this study, we identified CCN1 as a gene that is differentially up-regulated in alveolar mesenchymal cells of human subjects with rapidly progressive idiopathic pulmonary fibrosis (IPF). Elevated levels of CCN1 mRNA were confirmed in lung tissues of IPF subjects undergoing lung transplantation, and CCN1 protein was predominantly localized to fibroblastic foci. CCN1 expression in ex vivo IPF lung fibroblasts correlated with gene expression of the extracellular matrix proteins, collagen (Col)1a1, Col1a2, and fibronectin as well as the myofibroblast marker, a-smooth muscle actin. RNA interference (RNAi)-mediated knockdown of CCN1 down-regulated the constitutive expression of these profibrotic genes in IPF fibroblasts. TGF-b1, a known mediator of tissue fibrogenesis, induces gene and protein expression of CCN1 via a mothers against decapentaplegic homolog 3 (SMAD3)-dependent mechanism. Importantly, endogenous CCN1 potentiates TGF-b1-induced SMAD3 activation and induction of profibrotic genes, supporting a positive feedback loop leading to myofibroblast activation. In vivo RNAi-mediated silencing of CCN1 attenuates fibrogenic responses to bleomycin-induced lung injury. These studies support previously unrecognized, cooperative interaction between the CCN1 matricellular protein and canonical TGF-b1/SMAD3 signaling that promotes lung

show abstract

“…The phenotype of replicative senescence and stressinduced premature senescence has been likened by several groups to a form of post-mitotic differentiation (Rodemann et al 1991, Maas-Szabowski et al 1999, Alaluf et al 2000, Evans et al 2003. To evaluate the phenotype of the fibroblasts, samples were decoded and send for blind analysis to one of the authors (H. P. Rodemann).…”

Section: Phenotypic Analysis Of Fibroblastsmentioning

confidence: 99%