Abstract
Background: Our previous work demonstrated that application of transforming growth factor beta 1 (TGF-β1) and forskolin to the repair site after chronic denervation and axotomy has a mitogenic effect, reactivates Schwann cells (SCs), and supports axonal regeneration. We found decreased expression of fibroblast growth factor 7 (FGF-7), a factor involved in synaptic organization and maintenance. Using an in vitro system, we examined the molecular mechanism of TGF-β1 and forskolin on the regulation of FGF-7 expression in SCs. Methods: SCs were prepared from the sciatic nerve and stimulated with forskolin (0.5 μM), TGF-β1 (1 ng/mL), or TGF-β1 + forskolin for 6 or 24 hours. SCs were also pretreated with LY2109761 (0.5 μM), a TGF-β receptor inhibitor, prior to stimulation with TGF-β1 + forskolin for 6 hours. Real-time TaqMan quantitative polymerase chain reaction analyses for FGF-7, myelin basic protein, and peripheral myelin protein 22 expression were performed. Cycle threshold (Ct) data were normalized to a reference gene, and fold changes relative to untreated SCs were determined using the 2(-ΔΔCt) method. Statistical analysis was done using t test (P<0.05). Results: TGF-β1 alone or in combination with forskolin for 24 hours resulted in a 3.3- and 2.8-fold decrease in FGF-7 expression in SCs, respectively. No change in FGF-7 expression was found with forskolin alone. TGF-β1 + forskolin treatment for 6 hours resulted in a 4.0-fold decrease in FGF-7 expression, while the addition of LY2109761 resulted in a 2.7-fold decrease in FGF-7 expression. Conclusion: We showed that SC expression of FGF-7 is regulated by TGF-β1. The positive effect of TGF-β1 and forskolin on SC reactivation and axonal regeneration may involve modulation of FGF-7 expression and activity in SCs.