March 2012
Volume 53, Issue 14
ARVO Annual Meeting Abstract  |   March 2012
Changes In Bcl2, Bax, JNK, p65-NFkB And Smad7 Genes In Conjunctival Myofibroblasts Exposed To NGF
Author Affiliations & Notes
  • Alessandra Micera
    Lab Ophthalmology, IRCCS GB Bietti Eye Foundation, Rome, Italy
  • Bijorn O. Balzamino
    Lab Ophthalmology, IRCCS GB Bietti Eye Foundation, Rome, Italy
  • Filippo Biamonte
    Lab. Developmental Neuroscience and Neural Plasticity,
    University Campus BioMedico, Rome, Italy
  • Loredana Mastrella
    Dept. Ophthalmology,
    University Campus BioMedico, Rome, Italy
  • Stefano Bonini
    Dept. Ophthalmology,
    University Campus BioMedico, Rome, Italy
  • Francesca Levi-Schaffer
    Dept. Pharmacology, Institute for Drug Research, Hebrew University, Jerusalem, Israel
  • Footnotes
    Commercial Relationships  Alessandra Micera, None; Bijorn O. Balzamino, None; Filippo Biamonte, None; Loredana Mastrella, None; Stefano Bonini, None; Francesca Levi-Schaffer, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 3976. doi:
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      Alessandra Micera, Bijorn O. Balzamino, Filippo Biamonte, Loredana Mastrella, Stefano Bonini, Francesca Levi-Schaffer; Changes In Bcl2, Bax, JNK, p65-NFkB And Smad7 Genes In Conjunctival Myofibroblasts Exposed To NGF. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3976.

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      © ARVO (1962-2015); The Authors (2016-present)

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Purpose: : Nerve Growth Factor (NGF) might be involved in both healing and fibrotic process occurring in conjunctiva as a result of pathological conditions. We previously described the profibrogenic effect of NGF on conjunctival Fibroblasts, partially mediated by the release of Transforming Growth Factor-β1 (TGFβ1). The aim of this study was to quantify and compare caspase, cJun, p65-NFkB and smad7 expression and/or activation in NGF and TGFβ1 exposed myoFBs.

Methods: : Human conjunctival Fibroblast ( were expanded and TGFβ1-induced myofibroblast (myoFBs) were developed according to a stabilize setting. Serum-starved myoFBs were exposed to single/repeated increasing doses of NGF (0-200ng/mL) or TGFβ1 (10ng/mL) for 15min and 1, 3, 5, 24, 48 and 72 hrs. MTS and HO342 staining were performed. Pellets were analysed for Bcl2, Bax, smad7 and caspase3 by real-time PCR, FACS and confocal analysis. Both dosing and timing exposure results were statistically compared.

Results: : NGF effects on myoFBs are strongly dependent on expression ratio of trkANGFR/p75NTR. TGFβ1 by itself does not trigger significant p75NTR expression, retaining a high trkANGFR/p75NTR ratio. Further exposure to NGF shifted trkANGFR/p75NTR ratio in favour to p75NTR expression, with maximum effect at chronic NGF treatment. NGF triggered Bax expression, without affecting significantly Bcl2 expression. Smad7 was up-regulated in those myoFBs treated with NGF (p<.05). By contrary, TGFβ1 inhibited the pro-apoptotic activities and downregulated Bax expression (p<.05). Phosporylation of cJun and p65-NFkB translocation were detected as later as at chronic 100ng/mL NGF exposure.

Conclusions: : Together these findings suggest that NGF triggers apoptosis selectively in p75NTR-bearing myoFBs, mainly due to a selective p75NTR-dependent p65 translocation and JNK activation. The molecular mechanism behind this effect may also involve a deregulation of TGFβ1 signaling, as NGF exposure resulted also in Smad7 gene expression, a component of the TGFβ1 pathway with inhibitory activities.

Keywords: apoptosis/cell death • conjunctiva • growth factors/growth factor receptors 

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