Mechanisms involved in suppression of NGF-induced neuronal differentiation of PC12 cells by hyaluronic acid

In the present study, we found that hyaluronic acid (HA) suppressed the neuronal differentiation mediated by nerve growth factor (NGF). In addition, we examined the mechanism by which HA inhibits the NGF-induced neurite outgrowth of PC12 cells. We elucidated the direct interaction between NGF and HA, and found that HA did not bind to NGF directly using a quartz-crystal microbalance. Western blot analysis revealed that HA suppressed NGF-induced phosphorylation of p38 MAPK, ERKs, and transcriptional factor CREB in PC12 cells. Furthermore, HA inhibited the luciferase activity of pCRE-Luc transfected PC12 cells in the presence of NGF. We confirmed that the p38 MAPK inhibitor SB203580 and ERK inhibitor U0126 suppressed NGF-induced neurite outgrowth of PC12 cells, and found that the inhibitory effects of HA on phosphorylation of ERKs, but not of p38 MAPK, were restored by the anti-RHAMM antibody. The number of PC12 cells with neurites increased remarkably when pre-cultured with the anti-RHAMM antibody, then treated with NGF and HA. Our findings indicate that HA inhibits NGF-induced neuronal differentiation of PC12 cells partially by inhibiting ERK phosphorylation through RHAMM, and suggest that the binding of HA to RHAMM modifies the signaling pathways in PC12 cells treated with NGF.