Glycoprotein isolated from Cudrania tricuspidata Bureau inhibits iNO and COX-2 expression through modulation of NF-κB in LPS-stimulated RAW 264.7 cells

Glycoprotein of Cudrania tricuspidata Bureau (CTB glycoprotein) was isolated from CTB fruits which have been used to heal various disorders of the injury and lung as an herbal agent in Korea since long time ago. The CTB glycoprotein was identified to have a molecular weight of 75 kDa and consists of carbohydrate (72.5%) and protein moiety (27.5%). To know inhibitory ability of CTB glycoprotein for inflammation mediated by reactive oxygen radicals, firstly we tested about anti-oxidative activity (DPPH, superoxide anion, and hydroxyl radicals) in cell-free system, and then evaluated changes of inflammation-related signals [intracellular reactive oxygen species (iROS), nitric oxide (NO), nuclear factor-kappa B (NF-κB), COX-2, and iNOS] in the LPS (1 μg/ml)-treated RAW 264.7 cells. The results in this study showed that CTB glycoprotein (100 μg/ml) has a strong scavenging activity against DPPH, superoxide anion, and hydroxyl radicals without any pro-oxidant activity in vitro. In the inflammation-related signals, expression of iROS, NO, NF-κB, COX-2, and iNOS were inhibited by treatment with CTB glycoprotein (50 μg/ml) in the presence of LPS (1 μg/ml). Taken together, our data obtained from these experiments indicated that CTB glycoprotein suppresses expression of the inflammatory-related proteins (iNOS and COX-2) through regulation of NF-κB. Thus, we speculate that CTB glycoprotein may have therapeutic potential for inflammation-associated disorders.