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Endothelin and sex hormones modulate the fibronectin synthesis by cultured human skin scleroderma fibroblasts.

Soldano S, Montagna P, Villaggio B, Parodi A, Gianotti G, Sulli A, Seriolo B, Secchi ME, Cutolo M - Ann. Rheum. Dis. (2008)

Bottom Line: In normal FBs, ET-1 and 17beta-oestradiol, as well as their combination, increased cell growth (p<0.001, p<0.001, p<0.01 vs untreated cells (control), respectively) and fibronectin synthesis (p<0.05, p<0.05, p<0.01 vs control, respectively).By contrast, testosterone either alone or in combination with ET-1 did not influence cell proliferation, but decreased fibronectin synthesis (p<0.05, testosterone vs control).ET-1 and 17beta-oestradiol seem to exert a profibrotic effect in normal and SSc culture FBs and might suggest their synergistic effect in the pathogenesis of the fibrotic process in SSc.

View Article: PubMed Central - PubMed

Affiliation: Research Laboratories and Clinical Academic Unit of Rheumatology, Department of Internal Medicine, University of Genova, Italy.

ABSTRACT

Objective: To evaluate the influence of endothelin-1 (ET-1) and sex hormones on cell proliferation and extracellular matrix (ECM) synthesis (ie, fibronectin, laminin) by cultured normal and scleroderma (SSc) human skin fibroblasts (FBs).

Methods: Primary cultures of FBs were treated with ET-1 and sex hormones (17beta-oestradiol or testosterone) for 24 h. Cell growth was analysed by methiltetrazolium salt test, ECM synthesis was evaluated by immunocytochemistry and western blot, both at 24 h.

Results: In normal FBs, ET-1 and 17beta-oestradiol, as well as their combination, increased cell growth (p<0.001, p<0.001, p<0.01 vs untreated cells (control), respectively) and fibronectin synthesis (p<0.05, p<0.05, p<0.01 vs control, respectively). By contrast, testosterone either alone or in combination with ET-1 did not influence cell proliferation, but decreased fibronectin synthesis (p<0.05, testosterone vs control). In SSc FBs, ET-1 and 17beta-oestradiol alone or their combination induced an increased fibronectin synthesis (p<0.05, p<0.05, p<0.01 vs control, respectively). Unexpectedly, testosterone induced an increase of fibronectin synthesis (p<0.05 vs control).

Conclusions: ET-1 and 17beta-oestradiol seem to exert a profibrotic effect in normal and SSc culture FBs and might suggest their synergistic effect in the pathogenesis of the fibrotic process in SSc.

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Related in: MedlinePlus

A. Evaluation by 3-(4,5-dDimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test of cell proliferation in human normal fibroblasts (Fb) untreated (control) and treated for 24 h with endothelin-1 (ET-1) (10−7 M), 17β-oestradiol (E2) (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), testosterone (T) (10−9 M) or T (10−9 M) with ET-1 (10−7 M). **p<0.01; ***p<0.001. B. Evaluation by western blot analysis of proliferating cell nuclear antigen (PCNA) expression in human normal Fb untreated (control) and treated for 24 h with ET-1 (10−7 M), E2 (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), T (10−9 M) or T (10−9 M) with ET-1 (10−7 M). C. Evaluation by MTT test of cell proliferation in human systemic sclerosis fibroblasts (SSc Fb) untreated (control) and treated for 24 h with ET-1 (10−7 M), E2 (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), T (10−9 M) or T (10−9 M) with ET-1 (10−7 M).
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ard-68-04-0599-f01: A. Evaluation by 3-(4,5-dDimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test of cell proliferation in human normal fibroblasts (Fb) untreated (control) and treated for 24 h with endothelin-1 (ET-1) (10−7 M), 17β-oestradiol (E2) (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), testosterone (T) (10−9 M) or T (10−9 M) with ET-1 (10−7 M). **p<0.01; ***p<0.001. B. Evaluation by western blot analysis of proliferating cell nuclear antigen (PCNA) expression in human normal Fb untreated (control) and treated for 24 h with ET-1 (10−7 M), E2 (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), T (10−9 M) or T (10−9 M) with ET-1 (10−7 M). C. Evaluation by MTT test of cell proliferation in human systemic sclerosis fibroblasts (SSc Fb) untreated (control) and treated for 24 h with ET-1 (10−7 M), E2 (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), T (10−9 M) or T (10−9 M) with ET-1 (10−7 M).

Mentions: ET-1, E2 alone or ET-1 and E2 in combination induced a significant increase of cell proliferation (p<0.001; p<0.001; p<0.01, respectively), whereas T alone or in combination with ET-1 did not induced differences in normal human skin FBs when compared to the untreated controls (vs control) (fig 1A). The mean values between E2 and T differed significantly (p<0.001) (fig 1A). By contrast, T in combination with ET-1 decreased the cell growth (p<0.01) when compared to ET-1-treated FBs (fig 1A). These data were detected by the MTT test.


Endothelin and sex hormones modulate the fibronectin synthesis by cultured human skin scleroderma fibroblasts.

Soldano S, Montagna P, Villaggio B, Parodi A, Gianotti G, Sulli A, Seriolo B, Secchi ME, Cutolo M - Ann. Rheum. Dis. (2008)

A. Evaluation by 3-(4,5-dDimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test of cell proliferation in human normal fibroblasts (Fb) untreated (control) and treated for 24 h with endothelin-1 (ET-1) (10−7 M), 17β-oestradiol (E2) (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), testosterone (T) (10−9 M) or T (10−9 M) with ET-1 (10−7 M). **p<0.01; ***p<0.001. B. Evaluation by western blot analysis of proliferating cell nuclear antigen (PCNA) expression in human normal Fb untreated (control) and treated for 24 h with ET-1 (10−7 M), E2 (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), T (10−9 M) or T (10−9 M) with ET-1 (10−7 M). C. Evaluation by MTT test of cell proliferation in human systemic sclerosis fibroblasts (SSc Fb) untreated (control) and treated for 24 h with ET-1 (10−7 M), E2 (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), T (10−9 M) or T (10−9 M) with ET-1 (10−7 M).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2651484&req=5

ard-68-04-0599-f01: A. Evaluation by 3-(4,5-dDimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test of cell proliferation in human normal fibroblasts (Fb) untreated (control) and treated for 24 h with endothelin-1 (ET-1) (10−7 M), 17β-oestradiol (E2) (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), testosterone (T) (10−9 M) or T (10−9 M) with ET-1 (10−7 M). **p<0.01; ***p<0.001. B. Evaluation by western blot analysis of proliferating cell nuclear antigen (PCNA) expression in human normal Fb untreated (control) and treated for 24 h with ET-1 (10−7 M), E2 (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), T (10−9 M) or T (10−9 M) with ET-1 (10−7 M). C. Evaluation by MTT test of cell proliferation in human systemic sclerosis fibroblasts (SSc Fb) untreated (control) and treated for 24 h with ET-1 (10−7 M), E2 (10−10 M), E2 (10−10 M) with ET-1 (10−7 M), T (10−9 M) or T (10−9 M) with ET-1 (10−7 M).
Mentions: ET-1, E2 alone or ET-1 and E2 in combination induced a significant increase of cell proliferation (p<0.001; p<0.001; p<0.01, respectively), whereas T alone or in combination with ET-1 did not induced differences in normal human skin FBs when compared to the untreated controls (vs control) (fig 1A). The mean values between E2 and T differed significantly (p<0.001) (fig 1A). By contrast, T in combination with ET-1 decreased the cell growth (p<0.01) when compared to ET-1-treated FBs (fig 1A). These data were detected by the MTT test.

Bottom Line: In normal FBs, ET-1 and 17beta-oestradiol, as well as their combination, increased cell growth (p<0.001, p<0.001, p<0.01 vs untreated cells (control), respectively) and fibronectin synthesis (p<0.05, p<0.05, p<0.01 vs control, respectively).By contrast, testosterone either alone or in combination with ET-1 did not influence cell proliferation, but decreased fibronectin synthesis (p<0.05, testosterone vs control).ET-1 and 17beta-oestradiol seem to exert a profibrotic effect in normal and SSc culture FBs and might suggest their synergistic effect in the pathogenesis of the fibrotic process in SSc.

View Article: PubMed Central - PubMed

Affiliation: Research Laboratories and Clinical Academic Unit of Rheumatology, Department of Internal Medicine, University of Genova, Italy.

ABSTRACT

Objective: To evaluate the influence of endothelin-1 (ET-1) and sex hormones on cell proliferation and extracellular matrix (ECM) synthesis (ie, fibronectin, laminin) by cultured normal and scleroderma (SSc) human skin fibroblasts (FBs).

Methods: Primary cultures of FBs were treated with ET-1 and sex hormones (17beta-oestradiol or testosterone) for 24 h. Cell growth was analysed by methiltetrazolium salt test, ECM synthesis was evaluated by immunocytochemistry and western blot, both at 24 h.

Results: In normal FBs, ET-1 and 17beta-oestradiol, as well as their combination, increased cell growth (p<0.001, p<0.001, p<0.01 vs untreated cells (control), respectively) and fibronectin synthesis (p<0.05, p<0.05, p<0.01 vs control, respectively). By contrast, testosterone either alone or in combination with ET-1 did not influence cell proliferation, but decreased fibronectin synthesis (p<0.05, testosterone vs control). In SSc FBs, ET-1 and 17beta-oestradiol alone or their combination induced an increased fibronectin synthesis (p<0.05, p<0.05, p<0.01 vs control, respectively). Unexpectedly, testosterone induced an increase of fibronectin synthesis (p<0.05 vs control).

Conclusions: ET-1 and 17beta-oestradiol seem to exert a profibrotic effect in normal and SSc culture FBs and might suggest their synergistic effect in the pathogenesis of the fibrotic process in SSc.

Show MeSH
Related in: MedlinePlus