Limits...
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.

Show MeSH

Related in: MedlinePlus

Evaluation by immunocytochemistry (A) and western blot (B) of fibronectin (FN) expression 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). C. Evaluation by immunocytochemistry of laminin 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). Evaluation by immunocytochemistry (D) and western blot (E) of fibronectin expression in human systemic sclerosis (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). F. Evaluation by immunocytochemistry of laminin expression in human 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). *p<0.05, **p<0.01, ***p<0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

ard-68-04-0599-f02: Evaluation by immunocytochemistry (A) and western blot (B) of fibronectin (FN) expression 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). C. Evaluation by immunocytochemistry of laminin 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). Evaluation by immunocytochemistry (D) and western blot (E) of fibronectin expression in human systemic sclerosis (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). F. Evaluation by immunocytochemistry of laminin expression in human 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). *p<0.05, **p<0.01, ***p<0.001.

Mentions: ET-1 induced a significant increase of fibronectin synthesis (p<0.05 vs control), whereas no significant difference for laminin was found (fig 2A,C). E2 alone and in combination with ET-1 induced a further significant increase for fibronectin (p<0.05, p<0.01 vs control, respectively) and laminin (p<0.05, p<0.05 vs control, respectively) (fig 2A,C). Conversely, T was found to reduce the expression of both ECM proteins: the decrease was significant for fibronectin (p<0.05 vs control) in particular. The fibronectin synthesis was also significantly reduced in FBs cultured in the presence of T with ET-1 when compared to ET-1-treated cells (p<0.001) (fig 2A). Interestingly, the mean values between E2 and T differed significantly for fibronectin (p<0.001) and laminin (p<0.001) synthesis (fig 2A,C). These data were obtained by immunocytochemistry (ICC).


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)

Evaluation by immunocytochemistry (A) and western blot (B) of fibronectin (FN) expression 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). C. Evaluation by immunocytochemistry of laminin 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). Evaluation by immunocytochemistry (D) and western blot (E) of fibronectin expression in human systemic sclerosis (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). F. Evaluation by immunocytochemistry of laminin expression in human 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). *p<0.05, **p<0.01, ***p<0.001.
© 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-f02: Evaluation by immunocytochemistry (A) and western blot (B) of fibronectin (FN) expression 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). C. Evaluation by immunocytochemistry of laminin 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). Evaluation by immunocytochemistry (D) and western blot (E) of fibronectin expression in human systemic sclerosis (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). F. Evaluation by immunocytochemistry of laminin expression in human 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). *p<0.05, **p<0.01, ***p<0.001.
Mentions: ET-1 induced a significant increase of fibronectin synthesis (p<0.05 vs control), whereas no significant difference for laminin was found (fig 2A,C). E2 alone and in combination with ET-1 induced a further significant increase for fibronectin (p<0.05, p<0.01 vs control, respectively) and laminin (p<0.05, p<0.05 vs control, respectively) (fig 2A,C). Conversely, T was found to reduce the expression of both ECM proteins: the decrease was significant for fibronectin (p<0.05 vs control) in particular. The fibronectin synthesis was also significantly reduced in FBs cultured in the presence of T with ET-1 when compared to ET-1-treated cells (p<0.001) (fig 2A). Interestingly, the mean values between E2 and T differed significantly for fibronectin (p<0.001) and laminin (p<0.001) synthesis (fig 2A,C). These data were obtained by immunocytochemistry (ICC).

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