Glycoprotein isolated from Ulmus davidiana Nakai inhibits TPA-induced apoptosis through nuclear factor-kappa B in NIH/3T3 cells.

Glycoprotein of Ulmus davidiana Nakai (UDN glycoprotein) was isolated and identified using SDS-PAGE. UDN glycoprotein was shown to have strong scavenging activities against oxygen free radicals, as detected by different oxygen-radical formation assays. To investigate the anti-apoptotic effects of UDN glycoprotein, we investigated the activity of protein kinase C alpha (PKCalpha), the DNA-binding activation of nuclear factor-kappa B (NF-kappaB), the production of nitric oxide (NO) and apoptosis in 12-O-tetradecanoylphorbol 13-acetate (TPA)-stimulated NIH/3T3 cells using a western blot analysis, electrophoretic mobility shift assays (EMSA) and NO assays. Results in this experiment showed that 100 microg/ml of UDN glycoprotein has inhibitory effects on PKCalpha translocation, NF-kappaB DNA binding activity, NO production, and apoptosis in TPA (61.68 ng/ml)-stimulated NIH/3T3 cells. Interestingly however, it could not regulate the DNA binding activity of AP-1. Therefore, UDN glycoprotein, a natural anti-oxidant, is a potential modulator of apoptotic signal pathways in NIH/3T3 cells.