Thrombin stimulates production of fibronectin by human proximal tubular epithelial cells via a transforming growth factor- -dependent mechanism

Shirato, Kenichi; Osawa, Hiroshi; Kaizuka, Mitsuaki; Nakamura, Norio; Sugawara, Toshiyuki; Nakamura, Masayuki; Tamura, Michiko; Yamabe, Hideaki; Okumura, Ken

doi:10.1093/ndt/gfg398

Cited by 19 publications

(23 citation statements)

References 19 publications

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“…Such changes are similar to the promotion of matrix expression seen in other cells [10, 11, 13] and may contribute to the increased accumulation of interstitial collagen I found in progressive renal disease. Interestingly, the dose-response effect of thrombin on collagen mRNA expression was somewhat different to that seen with cell kinetics.…”

Section: Discussionsupporting

confidence: 55%

“…In this way, thrombin has been shown to regulate diverse functions in platelets, endothelial cells []reviewed in [4, 7], epithelial cells [8,9,10], and a variety of mesenchymal cells [11,12,13]. However, despite the functional significance of the interstitial fibroblast, the effects of thrombin on renal fibroblast function have not been determined.…”

Section: Introductionmentioning

confidence: 99%

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Thrombin Is a Pro-Fibrotic Factor for Rat Renal Fibroblasts in vitro

Hewitson

Martic²,

Kelynack³

et al. 2005

Nephron Exp Nephrol

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Background: Generation of thrombin occurs in response to parenchymal injury. Thrombin not only converts plasma fibrinogen into an insoluble fibrin clot, but also potentially augments inflammation through receptor-mediated activity. This study examines whether thrombin may potentially exacerbate fibrosis by upregulating the function of interstitial fibroblasts in vitro. Methods: Fibroblasts were isolated by explant outgrowth culture of rat kidneys. Subcultured cells were grown in DMEM+10% FCS supplemented with 0.1–0.5 U/ml thrombin. Functional parameters examined included kinetics (thymidine incorporation and change in cell number), differentiation (Western blotting for α-smooth muscle actin; αSMA), expression of procollagen α1(I) (Northern blotting) and contraction of collagen I lattices. RT-PCR was used to characterise expression of protease-activated receptors (PAR) previously implicated in thrombin’s cellular effects. Results: Cell population growth was increased 66 ± 41 and 47 ± 41% by 0.1 and 0.5 U/ml thrombin respectively (both p < 0.05 vs. basal). Likewise, 0.5 U/ml thrombin increased corrected procollagen α1(I) expression 2.4-fold (p < 0.05 vs. basal) and exacerbated the ability of fibroblasts to contract collagen matrix (p < 0.05 vs. basal). These effects were not associated with any change in expression of the myofibroblast marker αSMA. Effects on cell number were inhibited by treatment with (D)-Phe-Pro-Arg-chloromethylketone HCl (PPACK) suggesting that functional effects were mediated by serine protease activity. PAR-1 was the only fully functional known thrombin receptor expressed by these cells. Conclusion: Thrombin is a potential unrecognised fibroblast agonist in renal disease. Further studies of thrombin and its receptors may yield valuable insights into the pathogenesis of interstitial fibrosis.

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Section: Discussionsupporting

confidence: 55%

Section: Introductionmentioning

confidence: 99%

Thrombin Is a Pro-Fibrotic Factor for Rat Renal Fibroblasts in vitro

Hewitson

Martic²,

Kelynack³

et al. 2005

Nephron Exp Nephrol

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“…16) We have also reported that thrombin mediated various factors. [17][18][19][20][21][22] In the previous study, we showed that thrombin increased the production of MCP-1 and MIP-2 by GEC and enhanced mRNA of these cytokines. 23) It is well known that neutrophils are involved in acute phase of inflammation and following macrophages infiltrate.…”

Section: Discussionmentioning

confidence: 99%

Mizoribine Suppresses Proliferation of Rat Glomerular Epithelial Cells in Culture and Inhibits Increase of Monocyte Chemoattractant Protein-1 and Macrophage Inflammatory Protein-2 Stimulated by Thrombin

Yamabe

Shimada

Murakami

et al. 2012

Biological & Pharmaceutical Bulletin

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“…It can stimulate collagen synthesis in mesangial cells [28,29], and can enhance FN production by human proximal tubular epithelial cells [30]. In this study, the stimulatory effect of thrombin on MSCs was investigated.…”

Section: Introductionmentioning

confidence: 99%

Thrombin promotes fibronectin secretion by bone marrow mesenchymal stem cells via the protease-activated receptor mediated signalling pathways

Jin

Wang

et al. 2014

Stem Cell Res Ther

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IntroductionFibronectin (FN) is commonly used in the development of serum-free media for the expansion of mesenchymal stem cells (MSCs). This study was aimed to observe if thrombin could stimulate FN secretion by human bone marrow MSCs and investigate the potential underlying mechanisms.MethodsPCR was performed to detect the expression of the protease-activated receptors (PARs) in MSCs. After thrombin treatment, the expression level and secretion of FN were observed by RT-PCR, immunofluorescence staining and ELISA, respectively, and the activation of ERK1/2 and NF kappa B pathways was revealed by Western blotting, with or without pre-treatment of small-molecule blockers specific for PAR-1 and –2. The phenotypic and functional activities of thrombin-treated MSCs were also observed.ResultsPCR analysis showed that human bone marrow MSCs expressed two subtypes of PARs, PAR-1 and PAR-2. Thrombin treatment enhanced MSCs to express FN at mRNA and protein levels and promoted FN secretion by MSCs, accompanied by potent adherence to the culture plastic. Thrombin induced prompt phosphorylation of ERK 1/2 and NF kappa B p65 and the stimulatory effects of thrombin on FN secretion were blunted by specific inhibitors of these signaling molecules. Blockage to PAR-1 and PAR-2 partially abrogated thrombin-elicited FN secretion by MSCs and ERK 1/2 phosphorylation, whereas that of NF kappa B p65 was unaffected. Moreover, thrombin-treated MSCs maintained the phenotypic features, in vitro osteogenesis and adipogenesis capacities, and inhibitory activity on Phytohemagglutinin-induced allogeneic lymphocyte proliferation.ConclusionsThrombin could promote FN secretion by MSCs via PAR-mediated ERK 1/2 activation, while NF kappa B might be also involved in an undefined manner.

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Thrombin stimulates production of fibronectin by human proximal tubular epithelial cells via a transforming growth factor- -dependent mechanism

Cited by 19 publications

References 19 publications

Thrombin Is a Pro-Fibrotic Factor for Rat Renal Fibroblasts in vitro

Thrombin Is a Pro-Fibrotic Factor for Rat Renal Fibroblasts in vitro

Mizoribine Suppresses Proliferation of Rat Glomerular Epithelial Cells in Culture and Inhibits Increase of Monocyte Chemoattractant Protein-1 and Macrophage Inflammatory Protein-2 Stimulated by Thrombin

Thrombin promotes fibronectin secretion by bone marrow mesenchymal stem cells via the protease-activated receptor mediated signalling pathways

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