Nerve Growth Factor (NGF) modulates in vitro induced myofibroblasts by highlighting a differential protein signature

Abstract

We previously described the profibrogenic effect of NGF on conjunctival Fibroblasts (FBs) and its ability to trigger apoptosis in TGFβ1-induced myofibroblasts (myoFBs). Herein, cell apoptosis/signalling, cytokines’ signature in conditioned media and inflammatory as well as angiogenic pathway were investigated. Experimental myoFBs were exposed to NGF (0.1–100 ng/mL), at defined time-point for confocal and biomolecular analysis. Cells were analysed for apoptotic and cell signalling activation in cell extracts and for some inflammatory and proinflammatory/angiogenic factors’ activations. NGF triggered cJun overexpression and phospho-p65-NFkB nuclear translocation. A decreased Bcl2:Bax ratio and a significant expression of smad7 were confirmed in early AnnexinV-positive myoFBs. A specific protein signature characterised the conditioned media: a dose dependent decrease occurred for IL8, IL6 while a selective increase was observed for VEGF and cyr61 (protein/mRNA). TIMP1 levels were unaffected. Herein, NGF modulation of smad7, the specific IL8 and IL6 as well as VEGF and cyr61 modulation deserve more attention as opening to alternative approaches to counteract fibrosis.

Nerve Growth Factor (NGF) as Partaker in the Modulation of UV-Response in Cultured Human Conjunctival Fibroblasts

“…Most of the NGF effects on UV-exposed FBs occurred mainly at the lower NGF concentrations, suggesting the contribution of the high affinity trkA NGFR /p75 NTR receptor [ 11 , 26 , 52 ]. This aspect is not surprising, as encouraged by previous studies highlighting the survival role of the high-affinity NGF receptor in tissue homeostasis [ 28 , 36 ].…”

“…Highly involved in the homeostasis of the visual system, the nerve growth factor (NGF), the tyrosine kinase trkA NGF , and the pan-neurotrophin p75 NGF receptors have been recently reported to have multiple effects in UV-exposed tissues and cells [ 24 , 25 , 26 , 27 , 28 ]. Since NGF has been proven as (i), protective and therapeutic for damaged corneal and conjunctival epithelial cells; (ii), profibrogenic for corneal and conjunctival fibroblasts/myofibroblasts; and (iii), neuroprotective for retinal cells [ 10 , 11 ], the aim of this study was to explore the effect of NGF supplementation in UV-exposed conjunctival FBs by analyzing a few selected targets (inflammatory and epigenetic factors) previously linked to UV-exposed cells.…”